Novel anti-infective and anti-inflammatory compounds

ABSTRACT

Lysosomally accumulated substances that release a nitroxy group, or a short chain fatty acid or a product of anaerobic metabolism or a thiol or a sulfide often from an ester or similar labile linkage have anti-inflammatory, anti-cancer and anti-bacterial activity. They are useful in treating infectious, inflammatory and malignant disease.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.62/466,827, filed Mar. 3, 2017, which is incorporated herein byreference in its entirety.

BACKGROUND

The human body senses bacteria, fungi or other parasites by bothrecognition of their surface patterns and via reception of the metabolicproducts of those organisms. In particular, the immune system issensitive to the presence of anaerobic organisms which, by their natureare able to infect poorly perfused tissue, or emanate from the anaerobiclumen of the gut. Anaerobes seem to be a particular source of dangersignals for the immune system because they often produce toxins and areable to proliferate in the absence of oxygen, where immune cells areless active and less able to use oxidative burst to kill the bacteriathey ingest.

Amongst the signals that human cells respond to are the products offermentation such as the short chain fatty acids (SCFAs) acetate,propionate and butyrate. Less characterised mediators with similarfunction include: lactic acid, H₂S, HS—, nitrite, polyamines and similardecarboxylated amino acids such as 3-Indolepropionic acid (IPA),deoxybileacids and polyphenol metabolites like phenylpropionic acid. Ina most general sense, these metabolites are variously sensed by a rangeof receptors that include SCFA receptors (e.g. FFA2), the pregnane Xreceptor or the arylhydrocarbon receptor. For convenience, we willdescribe here these materials as Products of Anaerobic Metabolism (PAMor PAMS for the plural). Another type of signal is that of bacterialcell wall materials or bacterial nucleic acids. These materials areoften ligands for the Toll-like-receptor (TLR) family. These arereferred to as Pathogen-associated molecular patterns or PAMPs. Forsimplicity here, we will refer to all signal types as PAM or PAMs in theplural.

Although signaling of this type is commonly associated with the gutepithelium, it is also a potential signal in the interaction between theimmune system surveilling the gut or the periphery. In particular, thelysosomes of the gut or the immune system cells are exposed to bacterialmetabolites because these materials are released as bacteria are lysedfollowing phagocytosis.

We have observed that delivering donors of SCFAs, TLR ligands, and otherbacterial metabolites to the phagosome of immune cells results in immunestimulation such that phagocytosed bacteria are more rapidly killed.This effect can be augmented by the presentation of additional signalsin parallel, for example nitric oxide (NO), delivered from a nitroester.

Nitric oxide functions as a neurotransmitter, autacoid constitutivemediator, inducible mediator, cytoprotective molecule, and cyctotoxicmolecule. Since NO plays multiple physiological roles in the regulationof numerous and diverse organ functions, defects in the NO pathway leadto a variety of pathophysiological states. Possible disorders are:arterioscleroses, hypertension, coronary artery disease, cardiacfailure, pulmonary hypertension, stroke, impotence, gastrointestinalulcers, asthma, and other CNS and systemic disorders^([1]).

Synthetic chemical reagents that release NO continuously over a periodof time, under physiological conditions, have been in use for a longtime in treatment of cardiovascular diseases^([2]). Most widely used areorganic nitrates (e. g. glyceryl trinitrate): These NO donors needthiols as a cofactor for generating NO. They can use endogenous sourcesof thiols.

Other important series of NO donors are the so called NOC, NOR, andNONOate compounds which were reviewed by Wang et al^([3]).

Furthermore a class of activators of soluble guanylyl cyclase (e. g.YC-1) is known as NO-sensitizer, which may potentiate the effect ofminimal NO concentrations^([4]).

One major drawback of all those NO donors consists in the fact that theyexert their action largely in the extracellular environment. Forexample, nitro-glycerine leads to the release of NO in the plasma whichstimulates vasodilation by its action on smooth muscle surroundingvessels.

A similar drawback for the use of short chain fatty acids or hydrogensulfides as pharmaceutical agents is that they are potent odorants andrequired in relatively large amounts (the SCFA receptors have affinitiesin the milli-molar range). Thus they require specific delivery if theyare to be effective.

Another difficulty in the use of NO donors is the synthetic methods. Thepreparation of nitro and nitrooxy compounds belongs to the mostwidespread examples for electrophilic substitution reactions. In generalall methods for nitration lead to the formation of nitronium cations aselectrophiles, in most cases generated in situ^([5]). Few examples arepublished employing NO species as a salt, e.g. nitroniumtetrafluoroborate that can be employed for highly regioselectivearomatic nitration^([6]). Obstacles common to all methods are, besidesdesired selectivity, relatively harsh reaction conditions: acidity,oxidizing reactants, and temperature. Thus, classical procedures arelimited to stable systems that withstand these reaction conditions.Unfortunately this excludes many substance classes like drugs, organicmolecules, reducing sugars or other natural products. Transformation ofthese compounds to corresponding nitrates would result in valuablecompounds. Especially in the case of O-nitration synthesis faces severalproblems: Starting materials or intermediates in many cases do notwithstand conventional reaction conditions like HNO₃-H₂SO₄-mixtures.Thus the strategy of synthesis has to be changed, utilizing mildnitrating agents like acetyl nitrate^([7,8,9]) or benzoylnitrate^([9,10,11,12]), herein referred to as acyl nitrates. Chemicallytheses nitrating agents are mixed anhydrides from nitric acid andcorresponding carboxylic acids, mainly generated in situ by reaction ofa carboxylic anhydride with nitric acid.

NO has many biological functions and as such can serve as a molecularwarhead if appropriately delivered by a carrier molecule. In this regardit shares properties in common with compounds like CO and H₂S. Anotherclass of small effect molecule of natural origin are the short chainfatty acids (SCFAs) alluded to above. These compounds are products offermentation and in the gut serve as signals of microbial metabolismwhich are received by the gut epithelium and in turn used to coordinateanti-microbial homeostasis and epithelial microbial modulation.Similarly, TLR ligands are regulators of the immune response.

While all of these small natural modulators are known as extracellularsignals, there use as intracellular modulators is not described. Here wereport compounds that are designed to release these molecules in thecytoplasm and more importantly, acidic organelles such as the phagosomeor lysosome. Release of these molecules in the lysosome serves to informthe cell that it has digested a bacterium and thereby induces ananti-bacterial program that in turn enables a more robust response tointracellular organisms that may otherwise suppress bacteriolysis.

In particular we describe molecules which are acid trapped and able todonate a compound that is the product of anaerobic metabolism, Acidtrapped compounds are often amine containing compounds that areamphiphilic. They partition into the cell and concentrate in acidiccompartments due to their conversion to an ionized form at pH 5-6 whichis common in such organelles. Such acid-trapped molecules can beprepared with suitable linking groups such as hydroxyl groups. Multiplehydroxyl groups may be used to anchor one or more active molecules.There are many such acid-trapped compounds including common drugs suchas propranolol, amodiaquine, dextromethorphan, Dextrorphan, paroxetine,fluoxetine, astemizole or imipramine. Another example is the macrolideclass including compounds such as azithromycin, erythromycin orclarithromycin which are “acid trapped” in lysosomes by virtue of their2′ amine groups and amphilic properties. Azithromycin has two aminegroups and is particularly strongly trapped. These acid trappedmolecules can be derivatized, that is, decorated with signalingmolecules related to anaerobic metabolism such as SCFAs, NO, orHS-donors to form compounds of the invention. Using multiple signalingmolecules, or combinations thereof in multiple positions allows for aflexible means to tune the properties of the molecules. For example, wedescribed different effects for a compound carrying 3 SCFA esters versusone carrying a NO ester and a SCFA or a third donor and an SCFA. Inparticular, there is a hierarchy of effect with longer fatty acidspromoting differing immune responses. For example propionate differsfrom acetate in the degree of effect in this setting.

SUMMARY

The invention relates to compounds useful in modulating immune cellactivity or the barrier function of epithelial cells. The inventioncomprises compounds (e.g., derivative compounds of AmphiphilicLysosomally trapped Compounds (ALC), such ALC compounds including thoseof the formulae in any tables herein), which are subject to lysosomaltrapping and which bear moieties that are able to release TLR ligands,products of anaerobic metabolism, specifically SCFAs, sulfides,lactates, or NO, bile acids, polyamines, decarboxylated amino acids andpolyphenol metabolites like phenylpropionic acid.

The invention also provides a method of identifying a compound usefulfor modulating immune cell activity against bacteria: incubating such acompound with blood cells, preferably leukocytes, providing those cellswith bacteria, incubating the cells with bacteria, washing the cells andtreating them with a non-permeable antibiotic to reduce extracellularbacteria, then counting intracellular bacteria to observe whichcompounds reduce the number of intracellular bacteria surviving.

Optionally, it is advantageous to determine the ratio of theconcentration of the compound in the immune cells to non-immune cellssuch as erythrocytic cells as a measure of its lysosomal partition.Preferred are compounds that are preferentially taken up by immunecells.

In some embodiments the carrier compounds are macrolides with at leastone ONO₂-, SNO₂- or NNO₂-moiety. In other embodiments the carriercompounds are macrolides with at least one SCFA-moiety. In otherembodiments, the carrier molecule is amphiphilic with at least oneprotonatable amine. The term “macrolide” refers to any macrocycliclactone with 10 or more atoms connected within the ring system.Reference to an atom includes all isotopes of that atom. For example,structures drawn with carbon or hydrogen include isotopes such as ¹³C or²H.

An anti-microbial compound is a compound that inhibits the growth ordivision or replication of an organism such as a virus, bacteria,fungus, parasite, mycoplasma or other pathogen.

One embodiment is a compound comprising an Amphiphilic Lysosomallytrapped Compound (ALC) conjugated via an ester, thioester or nitroesterto a product of Anaerobic Metabolism (PAM) or one or more PAMs of thesame or different types. In a further embodiment, the compound is one inwhich the PAM is selected from one or more of Short Chain Fatty Acid(SCFA), NO, H₂S, mercaptans, polyamines, decarboxylated amino acids orpolyphenol metabolites like phenylpropionic acid. In a furtherembodiment, the compound is one in which the ALC is selected from amacrolide, polyamine, propranolol analog, chloroquine analog,amodiaquine, dextromethorphan, dextrorphan, paroxetine, fluoxetine,astemizole or imipramine analog.

Another embodiment is a macrolide comprising at least one ONO₂-, SNO₂-or NNO₂ moiety.

In some embodiments, the compound has the following formula (includingany possible salts thereof, except for nitrates, and any structures withexchanged isotopes, as possible by state of the art):

Formula 1

ALC conjugated or esterified with 1 or more of any of: X₍₁₋₅₎, Y₍₀₋₅₎,Z₍₀₋₃₎;

Where ALC=Amphiphilic Lysosomally trapped Compound;

X is a SCFA esterified to ALC and 0-5 indicates the number of moietiesconjugated;

Y is an NO donating group or an H₂S donating group esterified to ALC and0-5 indicates the number of moieties conjugated;

Z is a group donating sulfides, polyamines, decarboxylated amino acidsor polyphenol metabolites like phenylpropionic acid.

Wherein,

X=—N(CH₃)—CH₂—;

-   -   —CH₂—N(CH₃)—;    -   —C(═O)—;    -   —C(═NOR⁸)—;    -   —C(═NR¹²)—;

R¹ can be, but is not limited to

-   -   -(C₁-C₁₀)alkyl;    -   -(C₁-C₁₀)alkyliden-OH;    -   -(C₁-C₁₀)alkyliden-ONO₂;

R² can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷,        —C(═O)(NH)R⁷,—C(═S)(NH)R⁷;

R³ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,        —C(═S)(NH)R⁷;

If Z=O, R₄ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,        —C(═S)(NH)R⁷;

R₅ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,        —C(═S)(NH)R⁷;

or Z=O or NR⁹ and the R⁴ and R⁵ bearing atoms are connected via

-   -   —C(═O)— (If Z=O: carbonate linkage, if Z=NR⁹: carbamate linkage)    -   or the R⁴ and R⁵ bearing atoms are connected via W;        -   W may be but is not limited to        -   —(—)CH-(C₁-C₁₂)alkyl;        -   —(—)CH-(C₃-C₁₂)alkenyl;        -   —(—)CH-(C₃-C₁₂)alkynyl;        -   —(—)CH-(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;        -   —(—)CH-(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   wherein alkyl, alkenyl, alkynyl are optionally substituted by        one to five substituents selected independently from halogen (as        can be F, Cl, Br, I), (C₁-C₄)alkyl, (C₁-C₄)alkenyl,        (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl, (C₁-C₆)heterocycloalkyl,        (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy, hydroxy, nitro,        cyano, azido, mercapto, —NR¹⁴R¹⁵, R¹⁴C(═O)—, R¹⁴C(═O)O—,        R¹⁴OC(═O)O—, R¹⁴NHC(═O)—, R¹⁴C(═O)NH—, R¹⁴R¹⁵NC(═O)—,        R¹⁴OC(═O)—, and —xNO₂ with x=O;S;N;

R⁶ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,        —C(═S)(NH)R⁷;

R⁷ can be independently chosen from:

-   -   —H;    -   -ferrocene;    -   -C₁-C₁₀ alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl,        alkylheteroaryl wherein alkyl, alkenyl, alkynyl, aryl,        heteroaryl, alkylaryl and alkylheteroaryl groups are optionally        substituted by one to five substituents selected independently        from: ferrocene, halogen (as can be F, Cl, Br, I), (C₁-C₄)alkyl,        (C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,        (C₁-C₆)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl,        (C₁-C₄)alkoxy, hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido        (—N₃); mercapto (—SH), —NR¹⁴R¹⁵, R¹⁴C(═O)—, R¹⁴C(═O)O—,        R¹⁴OC(═O)O—, R¹⁴NHC(═O)—, R¹⁴C(═O)NH—, R¹⁴R¹⁵NC(═O)—,        R¹⁴OC(═O)—, and —XNO_((y)) with X=O; S; N and y=1 or 2;

R⁸ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)—R⁷;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;

R⁹ can be, but is not limited to

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)—R⁷;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;

R¹² can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)—R⁷;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;

R¹⁴, R¹⁵ can independently be, but are not limited to:

-   -   —H;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;    -   wherein alkyl, alkenyl, alkynyl, aryl and heteroaryl are        optionally substituted by one to five substituents selected        independently from halogen (as can be F, Cl, Br, I),        (C₁-C₄)alkyl, (C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,        (C₁-C₆)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl,        (C₁-C₄)alkoxy, hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido        (—N₃), mercapto (—SH), and —XNO_(y) with X=O; S; N and y=1 or 2;    -   or N(R¹⁴R¹⁵) is an aziridine, azetidine, pyrrolidine,        piperidine, azepane or azocane, 1-substituted piperazine, or        morpholine moiety.

In some embodiments, the compound has the following formula (includingany possible salts thereof, except for nitrates, and any structures withexchanged isotopes, as possible by state of the art):

Formula 1

ALC conjugated or esterified with 1 or more of any of: X₍₁₋₅₎, Y₍₀₋₅₎,Z₍₀₋₃₎;

Where ALC=Amphiphilic Lysosomally trapped Compound;

X is a SCFA esterified to ALC and 0-5 indicates the number of moietiesconjugated;

Y is an NO donating group or an H₂S donating group esterified to ALC and0-5 indicates the number of moieties conjugated;

Z is a group donating sulfides, polyamines, decarboxylated amino acidsor polyphenol metabolites like phenylpropionic acid.

Wherein,

X=—N(CH₃)—CH₂—;

-   -   —CH₂—N[(CH₂)_(n)CH₃]—; wherein n is 0-4;    -   —C(═O)—;    -   —C(═NOR⁸)—;    -   —C(═NR¹²)—;

R¹ can be, but is not limited to

-   -   -(C₁-C₁₀)alkyl;    -   -(C₁-C₁₀)alkyliden-OH;    -   -(C₁-C₁₀)alkyliden-ONO₂;

R² can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷,        —C(═O)(NH)R⁷,—C(═S)(NH)R⁷;

R³ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,        —C(═S)(NH)R⁷;

If Z=O, R⁴ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷,        —C(═O)(NH)R⁷,—C(═S)(NH)R⁷;

R⁵ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷,        —C(═O)(NH)R⁷,—C(═S)(NH)R⁷;

or Z=O or NR⁹ and the R⁴ and R⁵ bearing atoms are connected via

-   -   —C(═O)— (If Z=O: carbonate linkage, if Z=NR⁹: carbamate linkage)    -   or the R⁴ and R⁵ bearing atoms are connected via W;        -   W may be but is not limited to        -   —(—)CH-(C₁-C₁₂)alkyl;        -   —(—)CH-(C₃-C₁₂)alkenyl;        -   —(—)CH-(C₃-C₁₂)alkynyl;        -   —(—)CH-(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;        -   —(—)CH-(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   wherein alkyl, alkenyl, alkynyl are optionally substituted by        one to five substituents selected independently from halogen (as        can be F, Cl, Br, I), (C₁-C₄)alkyl, (C₁-C₄)alkenyl,        (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl, (C₁-C₆)heterocycloalkyl,        (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy, hydroxy, nitro,        cyano, azido, mercapto, —NR¹⁴R¹⁵, R¹⁴C(═O)—, R¹⁴C(═O)O—,        R¹⁴OC(═O)O—, R¹⁴NHC(═O)—, R¹⁴C(═O)NH—, R¹⁴R¹⁵NC(═O)—,        R¹⁴OC(═O)—, and —xNO₂ with x=O;S;N;

R⁶ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,        —C(═S)(NH)R⁷;

R⁷ can be independently chosen from:

-   -   —H;    -   -ferrocene;    -   -C₁-C₁₀ alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl,        alkylheteroaryl wherein alkyl, alkenyl, alkynyl, aryl,        heteroaryl, alkylaryl and alkylheteroaryl groups are optionally        substituted by one to five substituents selected independently        from: ferrocene, halogen (as can be F, Cl, Br, I), (C₁-C₄)alkyl,        (C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,        (C₁-C₆)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl,        (C₁-C₄)alkoxy, hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido        (—N₃); mercapto (—SH), —NR¹⁴R¹⁵, R¹⁴C(═O)—, R¹⁴C(═O)O—,        R¹⁴OC(═O)O—, R¹⁴NHC(═O)—, R¹⁴C(═O)NH—, R¹⁴R¹⁵NC(═O)—,        R¹⁴OC(═O)—, and —XNO_((y)) with X=O; S; N and y=1 or 2;

R⁸ can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)—R⁷;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;

R⁹ can be, but is not limited to

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)—R⁷;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;

R¹² can be, but is not limited to:

-   -   —H;    -   —NO_((y)) with y=1 or 2;    -   —C(═O)—R⁷;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;

R¹⁴, R¹⁵ can independently be, but are not limited to:

-   -   —H;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)alkynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;    -   wherein alkyl, alkenyl, alkynyl, aryl and heteroaryl are        optionally substituted by one to five substituents selected        independently from halogen (as can be F, Cl, Br, I),        (C₁-C₄)alkyl, (C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,        (C₁-C₆)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl,        (C₁-C₄)alkoxy, hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido        (—N₃), mercapto (—SH), and —XNO_(y) with X=O; S; N and y=1 or 2;    -   or N(R¹⁴R¹⁵) is an aziridine, azetidine, pyrrolidine,        piperidine, azepane or azocane, 1-substituted piperazine, or        morpholine moiety.

In some embodiments, the compound has the following formula (includingany any possible salts thereof, except for nitrates, and any structureswith exchanged isotopes, as possible by state of the art):

Wherein R¹, R², R⁴, R⁵, R⁶, X, and Z are defined as in formula 2;

R^(3a), R^(3b)=both —H;

-   -   or in the case R^(3a) is —H, R^(3b) can be:    -   —OH;    -   —OR¹⁴;    -   —NR¹⁴R¹⁵;    -   —C(═O)—R⁷;

or R^(3a)=R^(3b)=(═O);

-   -   =any possible cyclic or non-cyclic acetal;    -   (=NR¹²);    -   =any possible cyclic or non-cyclic aminal;    -   —OC(═O)R⁷;    -   OR¹⁴;

and R⁷, R¹⁴, and R¹⁵ are defined as in formula 2.

Where X can be O or S;

When X=O, R₁ may be but not limited to —(C═O)CH₃, —(C═O)CH₂CH₃,—(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, -13 (C═O)CHCHCOOH,—(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₂Y, or —(C═O)CH(ONO₂)CH₃;

R₂=R₃=H;

Y=can be a 5-membered saturated ring containing a disulfide bond;

When X=O, R₁ may be but not limited to —(C═O)CH₃, —(C═O)CH₂CH₃,—(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, —(C═O)CHCHCOOH,—(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₂Y, or —(C═O)CH(ONO₂)CH₃;

R₂=CH₃; R₃=H; or

When X=O, R₁=NO₂; R₂ consists of linker —CH₂CH₂OR₄, where R₄ may be butnot limited to —(C═O)CH₃, —(C═O)CH₂CH₃, —(C═O)CH₂CH₂CH₃,—(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH₃,—(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂, or—(C═O)CH₂CH₂CH₂CH₂Y; R3=H;

Y=can be a 5-membered saturated ring containing a disulfide bond; or

When X=O, R₁ may be but not limited to —(C═O)CH₃, —(C═O)CH₂CH₃,—(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, —(C═O)CHCHCOOH,—(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₂Y, or —(C═O)CH(ONO₂)CH₃;

Y=can be a 5-membered saturated ring containing a disulfide bond;

R² consists of linker —CH₂CH₂OR₄, where R₄=NO₂; R₃=H; or

When X=O, R₁ is NO₂, R₂=H or CH₃, R₃=OR₅, where R₅ may be but notlimited to —(C═O)CH₃, —(C═O)CH₂CH₃, —(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH,—(C═O)(C═O)CH₃, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₂Y, or—(C═O)CH(ONO₂)CH₃;

Y=can be a 5-membered saturated ring containing a disulfide bond; or

When X=S, R¹ may be but not limited to —(C═O)CH₃, a metal salt, or formsa disulfide bridge with itself, R²=R³=H.

Definition of Substituents on ALC Formulae (e.g., Macrofides,Hydroxychloroquine, Propranolol, etc.)

—NO₂ Nitro —(C═O)—C₆H₄—NO₂ p-nitrobenzoyl —(C═O)C₆H₅ benzoyl—(C═O)CH₂CH₂COOH succinyl —(C═O)CH₂CH₃ propionyl —(C═O)CH₂CH₂CH₃ butyryl—(C═O)CH₃ acetyl —(C═O)(C═O)CH₃ Pyruvyl —(C═O)CHCHCOOH Maleyl—(C═O)CH(OH)CH₃ Lactyl —(C═O)CH(ONO₂)CH₃ 2-O-Nitrolactyl —(C═O)C(CH₃)₂Isobutyryl —(C═O)CH₂CH₂CH₂CH₃ Valeryl —(C═O)CH₂C(CH₃)₂ Isovalericyl—(C═O)CH(CH₃)O(C═O)CH₃ Acetoxypropionyl —(C═O)CH₂CH₂(C═O)—ZSuccinyl-dithiole-3-thione —(C═O)OCH₂CH₂S(S)_(n)H Polysulfide ethylcarbonate —(C═O)OCH₂CH₂SNO NO-thioethylcarbonate —(C═S)OC₆H₅O-Phenylchlorothiono carbonate —(C═O)CH₂CH₂CH₂CH₂CH₃ hexanoyl —CH₂CH₂Brbromoethyl —(C═O)OCH₂CHCH₂ Vinyl carbonate —(C═O)CH₂CH₂CH₂CH₂Y Lipoyl

Wherein

Mac=a macrolide ring or macrolide ring system, for example, but notlimited to azithromycin or gamithromycin, each without the desosaminresidue.

Compounds with the Structure

Wherein

Mac=a macrolide ring or macrolide ring system, for example, but notlimited to azithromycin or gamithromycin, each without the desosaminresidue;

R″=independently of each other

—H;

—NO_((y)) with y=1 or 2;

—C(═O)OR³, —C(═S)OR³, —C(═O)R³, —C(═S)R³, —C(═O)(NH)R³, —C(=S)(NH)R³;

R¹, R₂=independently of each other H, OH, OR⁴, -C₁-C₁₀ alkyl, alkenyl,alkynyl, aryl, heteroaryl, alkylaryl, alkylheteroaryl;

wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl andalkylheteroaryl groups are optionally substituted by one to fivesubstituents selected independently from: fluorine, (C₁-C₄)alkyl,(C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,(C₁-C₉)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy,hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido (—N₃), mercapto (—SH),(C₁-C₄)alkthio, —NR⁴R⁵, R⁴C(═O)—, R⁴C(═O)O—, R⁴OC(═O)O—, R⁴NHC(═O)—,R⁴C(═O)NH—, R⁴R⁵NC(═O)—, R⁴OC(═O)— and —XNO_((y)) with X=O; S; N and y=1or 2;

or N(R₁R²) is an aziridine, azetidine, pyrrolidine, piperidine, azepaneor azocane, 1-substituted piperazine, or morpholine moiety;

R³=-C₁-C₁₀ alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl,alkylheteroaryl wherein alkyl, alkenyl, alkynyl, aryl, heteroaryl,alkylaryl and alkylheteroaryl groups are optionally substituted by oneto five substituents selected independently from: halogen, (C₁-C₄)alkyl,(C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,(C₁-C₉)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy,hydroxyl (—OH), nitro (—NO₂), cyano (—to CN), azido (—N₃), mercapto(—SH), (C₁-C₄)alkthio, —NR⁶R⁷, R⁶C(═O)—, R⁶C(═O)O—, R⁶OC(═O)O—,R⁶NHC(═O)—, R⁶C(═O)NH—, R⁶R⁷NC(═O)—, R⁶OC(═O)—, and —XNO_((y)) with X=O;S; N and y=1 or 2;

R⁴, R⁵, R⁶ and R⁷ can independently be, but are not limited to:

-   -   —H;    -   -(C₁-C₁₂)alkyl;    -   -(C₁-C₁₂)alkenyl;    -   -(C₁-C₁₂)akynyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;    -   -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;    -   -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;    -   -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl;        -   wherein alkyl, alkenyl, alkynyl, aryl and heteroaryl are            optionally substituted by one to five substituents selected            independently from ferrocene, halogen (as can be F, Cl, Br,            I), (C₁-C₄)alkyl, (C₁-C₄)alkenyl, (C₁-C₄)alkynyl,            (C₃-C₇)cycloalkyl, (C₁-C₉)heterocycloalkyl, (C₆-C₁₀)aryl,            (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy, hydroxyl (—OH),            (C₁-C₆)acyloxy, nitro (—NO₂), cyano (—CN), azido (—N₃),            mercapto (—SH), and —XNO_(y) with X=O; S; N and y=1 or 2.

Unless, otherwise stated, the word “macrolide”herein refers to thefamily of well-known macrolactone antibiotics and also the variousmacrolactone ALCs described herein.

In one aspect, the invention provides a composition comprising acompound of any of the formulae herein (e.g., any of the formulae, anyformula in the tables herein), or salt, solvate, hydrate or prodrugthereof, and a pharmaceutically acceptable carrier. In a further aspect,the composition can further comprise an additional therapeutic agent.

In one aspect, the invention provides a method of treating a subjectsuffering from or susceptible to a disease, disorder, or symptomthereof. The method includes administering to a subject in need thereofa therapeutically effective amount of a compound of any of the formulaeherein (e.g., any of the formulae, any formula in the tables herein), orsalt, solvate, hydrate or prodrug thereof. The disease, disorder, orsymptom thereof can be, for example, an infectious disease, aninflammatory disease, a malignant disease, a bacterial infection, aninflammatory reaction to a bacterial translocation event, aninflammation of the GI tract including intestines, colon, liver andpancreas, an inflammation of the airways, a systemic inflammatorydisease or a malignant or neoplastic disease.

In one aspect, the invention provides a method of stimulating immune orepithelial cells to form an anti-infective barrier or anti-infectiveresponse comprising contacting the cells with a compound of any of theformulae herein (e.g., any of the formulae, any formula in the tablesherein), or salt, solvate, hydrate or prodrug thereof. A further aspectof the method is wherein the contacting results in the intracellularrelease of a PAM comprising one or more of a molecule type selected fromSCFA, NO, H₂S, sulfides, polyamines, decarboxylated amino acids orpolyphenol metabolites like phenylpropionic acid from the compound ofany of the formulae herein (e.g., any of the formulae, any formula inthe tables herein), or salt, solvate, hydrate or prodrug thereof. Afurther aspect of the method is that comprising the intracellularrelease of one or more types of a short chain fatty acid moiety from thecompound of any of the formulae herein (e.g., any of the formulae, anyformula in the tables herein), or salt, solvate, hydrate or prodrugthereof. A further aspect of the method is that comprising theintracellular release of a short chain fatty acid moiety containing 2 ormore carbons from an appropriate carrier molecule.

In an embodiment, the compound of the invention is administered to thesubject using a pharmaceutically-acceptable formulation, e.g., apharmaceutically-acceptable formulation that provides sustained deliveryof the compound of the invention to a subject for at least 12 hours, 24hours, 36 hours, 48 hours, one week, two weeks, three weeks, or fourweeks after the pharmaceutically-acceptable formulation is administeredto the subject.

In certain embodiments, these pharmaceutical compositions are suitablebar topical or oral administration to a subject. In other embodiments,as described in detail below, the pharmaceutical compositions of thepresent invention may be specially formulated for administration insolid or liquid form, including those adapted for the following: (1)oral administration, for example, drenches (aqueous or non-aqueoussolutions or suspensions), tablets, boluses, powders, granules, pastes;(2) parenteral administration, for example, by subcutaneous,intramuscular or intravenous injection as, for example, a sterilesolution or suspension; (3) topical application, for example, as acream, ointment or spray applied to the skin; (4) intravaginally orintrarectally, for example, as a pessary, cream or foam; or (5) aerosol,for example, as an aqueous aerosol, liposomal preparation or solidparticles containing the compound.

The phrase “pharmaceutically acceptable” refers to those compound of theinventions of the present invention, compositions containing suchcompounds, and/or dosage forms which are, within the scope of soundmedical judgment, suitable for use in contact with the tissues of humanbeings and animals without excessive toxicity, irritation, allergicresponse, or other problem or complication, commensurate with areasonable benefit/risk ratio.

The term “pharmaceutically acceptable salts” or “pharmaceuticallyacceptable. carrier” is meant to include salts of the active compoundswhich are prepared with relatively nontoxic acids or bases, depending onthe particular substituents found on the compounds described herein.When compounds of the present invention contain relatively acidicfunctionalities, base addition salts can be obtained by contacting theneutral form of such compounds with a sufficient amount of the desiredbase, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable base addition salts include sodium,potassium, calcium, ammonium, organic amino, or magnesium salt, or asimilar salt. When compounds of the present invention contain relativelybasic functionalities, acid addition salts can be obtained by contactingthe neutral form of such compounds with a sufficient amount of thedesired acid, either neat or in a suitable inert solvent. Examples ofpharmaceutically acceptable acid addition salts include those derivedfrom inorganic acids like hydrochloric, hydrobromic, nitric, carbonic,monohydrogencarbonic, phosphoric, monohydrogenphosphoric,dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydroiodic, orphosphorous acids and the like, as well as the salts derived fromrelatively nontoxic organic acids like acetic, propionic, isobutyric,maleic, malonic, benzoic, succinic, suberic, fumaric, lactic, mandelic,phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric,methanesulfonic, and the like. Also included are salts of amino acidssuch as arginate and the like, and salts of organic acids likeglucuronic or galactunoric acids and the like (see, e.g., Berge et al.,Journal of Pharmaceutical Science 66:1-19 (1977)). Certain specificcompounds of the present invention contain both basic and acidicfunctionalities that allow the compounds to be converted into eitherbase or acid addition salts. Other pharmaceutically acceptable carriersknown to those of skill in the art are suitable for the presentinvention.

Some examples of substances which can serve as pharmaceutical carriersare sugars, such as lactose, glucose and sucrose; starches such as cornstarch and potato starch; cellulose and its derivatives such as sodiumcarboxymethycellulose, ethylcellulose and cellulose acetates; powderedtragancanth; malt; gelatin; talc; stearic acids; magnesium stearate;calcium sulfate; vegetable oils, such as peanut oils, cotton seed oil,sesame oil, olive oil, corn oil and oil of theobroma; polyols such aspropylene glycol, glycerine, sorbitol, mannitol, and polyethyleneglycol; agar; alginic acids; pyrogen-free water; isotonic saline; andphosphate buffer solution; skim milk powder; as well as other non-toxiccompatible substances used in pharmaceutical formulations such asVitamin C, estrogen and echinacea, for example. Wetting agents andlubricants such as sodium lauryl sulfate, as well as coloring agents,flavoring agents, lubricants, excipients, tableting agents, stabilizers,anti-oxidants and preservatives, can also be present. Solubilizingagents, including for example, cremaphore and beta-cyclodextrins canalso be used in the pharmaceutical compositions herein.

The neutral forms of the compounds may be regenerated by contacting thesalt with a base or acid and isolating the parent compound in theconventional manner. The parent form of the compound differs from thevarious salt forms in certain physical properties, such as solubility inpolar solvents, but otherwise the salts are equivalent to the parentform of the compound for the purposes of the present invention.

Initial dosages also can be estimated from in vivo data, such as animalmodels. Animal models useful for testing the efficacy of compounds totreat or prevent the various diseases described above are well-known inthe art.

Dosage amounts will typically be in the range of from about 0.0001 or0.001 or 0.01 mg/kg/day to about 100 mg/kg/day, (e.g., 0.01 to 1 mg/kgeffective dose) but can be higher or lower, depending upon, among otherfactors, the activity of the compound, its bioavailability, the mode ofadministration, and various factors discussed above. Dosage amount andinterval can be adjusted individually to provide plasma levels of thecompound(s) which are sufficient to maintain therapeutic or prophylacticeffect. In cases of local administration or selective uptake, such aslocal topical administration, the effective local concentration ofactive compound(s) cannot be related to plasma concentration. Skilledartisans will be able to optimize effective local dosages without undueexperimentation

The compound(s) can be administered once per day, a few or several timesper day, or even multiple times per day, depending upon, among otherthings, the indication being treated and the judgment of the prescribingphysician.

Preferably, the compound(s) will provide therapeutic or prophylacticbenefit without causing substantial toxicity. Toxicity of thecompound(s) can be determined using standard pharmaceutical procedures.The dose ratio between toxic and therapeutic (or prophylactic) effect isthe therapeutic index. Compounds(s) that exhibit high therapeuticindices are preferred.

The recitation of a listing of chemical groups in any definition of avariable herein includes definitions of that variable as any singlegroup or combination of listed groups. The recitation of an embodimentfor a variable herein includes that embodiment as any single embodimentor in combination with any other embodiments or portions thereof. Therecitation of an embodiment herein includes that embodiment as anysingle embodiment or in combination with any other embodiments orportions thereof.

Another object of the present invention is the use of a compound asdescribed herein (e.g., of any formulae herein) in the manufacture of amedicament for use in the treatment of a disorder or disease herein.Another object of the present invention is the use of a compound asdescribed herein (e.g., of any formulae herein) for use in the treatmentof a disorder or disease herein.

Many compounds of this invention have one or more double bonds, or oneor more asymmetric centers. Such compounds can occur as racemates,racemic mixtures, single enantiomers, individual diastereomers,diastereomeric mixtures, and cis- or trans- or E- or Z-double isomericforms.

Further, the aforementioned compounds also include their N-oxides. Theterm “N-oxides” refers to one or more nitrogen atoms, when present in acompound, are in N-oxide form, i.e., N→O.

Combinations of substituents and variables envisioned by this inventionare only those that result in the formation of stable compounds. Theterm “stable”, as used herein, refers to compounds which possessstability sufficient to allow manufacture and which maintains theintegrity of the compound for a sufficient period of time to be usefulfor the purposes detailed herein (e.g., treating a disease).

In another aspect the present invention relates to a mild and highlyselective process for the in situ introduction of the O—, S— andN-nitrate group into compounds of any othe formulae herein.

DETAILED DESCRIPTION OF THE INVENTION Technical Problem

NO, SCFA or PAM are difficult to use directly in Pharmaceuticalcompositions.

The use of NO donors is well known in medicine as a means to modulateblood pressure and inflammation^([13]). Such NO donors exert theiraction, as aforementioned, largely in the extracellular environment. Inthe case of blood pressure regulators, release of NO in the bloodresults in effects on the endothelium which are desirable. In the caseof modulators of inflammation, general release of NO may result in sideeffects such as loss of blood pressure which are undesirable.

Similarly, the SCFAs are volatile. malodourous and unstable. PAMs suchas polyamines are similarly unattractive for direct use. While theseproducts such as SCFAs or NO (inhaled gas) have been applied to thebody, they are used in amounts that are undesirable. The technicalproblem to solve, was, therefore, to focus the release of these products(NO, SCFA, PAMs) to cells associated with inflammation, cancer orinfection such that lower amounts could be used.

Solution to Problem

The compounds reported here are mostly located in intracellularcompartments and thus may donate NO or SCFA or PAM to an intracellularreceptor, preferably an intra-phagosomal or lysosomal receptor.Macrolide anti-bacterial compounds are well known for their ability tobe concentrated in acidic compartments, notably the phagosomes of immunecells such as neutrophils and macrophage^([14]). Phagosomes are theorgans where bacteria and other debris are digested by the phagocytesusing oxidative processes and digestive enzymes. Certain bacteria resistthis process by reducing the capacity of the cell to produceantibacterial factors (lower pH, proteases, active oxygen species,antibacterial enzymes, NO).

If, however, a compound was also trapped in the phagosome that wascapable of donating a stimulatory factor such as SCFA, PAM or NO, thenthere is the potential to overcome the inhibition due to the bacterium.More importantly, if the Phagocyte absorbs a compound able to donateSCFA, PAM or NO prior to that phagocyte encountering bacteria, it ispotentially stimulated to better kill bacteria immediately on contactwith them. This is potentially of significance in treating infections bybacteria such as Legionella, Pasteurella, Listeria and Mycobacteriumspecies that are intracellular parasites. It is also potentiallysignificant in the stimulation of barrier cells to resist the effect ofbacteria, or to maintain physical barriers toward bacteria.

In addition to their roles in immunology, SCFA, NO and PAM have a rolein homeostasis, acute inflammation and wound healing^([15]). Phagocyteslike macrophages are involved in many aspects of metabolism and aresensitive to SCFA, NO and PAM. The delivery of these substancespreferentially to cells of this type is a means to allow them to respondto the stimulus of SCFA, PAM or NO without using high systemic levels.This is achieved by delivering the substances as conjugates tolysosomally tropic compounds (ALCs).

Thus the efficacy of molecules described herein in various models ofinflammation and resolution of inflammation were examined. In thesemodels, example compounds reported here were able to reduce the effectsof inflammation, support body weight maintenance, and reduce diseasesigns without causing appreciable toxicity.

Definitions

“Anti-infective barrier” means the ability of epithelium to prevent thepenetration of bacteria or other pathogens.

Stimulating the formation of an anti-infective barrier means increasingthe ability of epithelium to prevent the penetration of bacteria orother pathogens through the up-regulation of tight junction formation orother barrier functions.

“Anti-infective response” means the ability of immune cells or similarto prevent the growth of bacteria or other pathogens via phagocytosis,oxidative burst or other toxic responses inactivating the pathogen.

Stimulating the formation of an anti-infective response means increasingthe ability of immune cells or similar cells to prevent the growth ofbacteria or other pathogens via phagocytosis, oxidative burst or othertoxic responses inactivating the pathogen.

“ALC” means an Amphiphilic Lysosomally trapped Compound.

“PAM” means Product of Anaerobic Metabolism. PAMs include but are notlimited to SCFA, NO, H₂S, mercaptans that eventually generate H₂S/HS⁻polyamines (e.g., compounds of Table 7), amino acid residues lacking theC-terminus (decarboxlated), bile acids (e.g., steroid acids found in thebile of mammals and other vertebrates, such as chenodeoxycholic acid,cholic acid, deoxycholic acid, lithocholic acid, and the like), ordegradation products from polyphenol metabolism such as3-(3-Hydroxyphenyl)propanoic (hMPP) acid, SCFA means Short Chain FattyAcid, which is a fatty acid molecule haying an aliphatic tail of eightor less carbon atoms.

“NO” means Nitric Oxide.

Advantageous Effects of Invention

The compounds reported here are useful in many respects. They areanti-microbial, anti-inflammatory, able to accumulate in tumors anddonate NO and able to protect against inflammation of the intestine.Selected embodiments are able to modulate inflammation of the liver andprotect against accumulation of fat or the resulting fibrosis.

The compounds are readily soluble as salts, may be provided by the oralroute, or via other means. They are adequately stable forpharmacological use when stored at the appropriate pH conditions.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 : TNFa-Production by LPS treated mice with either Vehicle (1%citric acid in water, 5 mL/kg) or 10 μmol/kg Compound E2 or Compound E3.

FIG. 2 : Change in body weight in mice in which arthritis has beeninduced using bovine collagen. Animals were treated with either Vehicle(1% citric acid in water, 5 mL/kg) or the indicated doses of compound E2μmol/kg. Data are from 10 animals per group, and data points significantdifferent from Vehicle are marked with *.

FIG. 3 : Number of mice maintaining body weight in which intestinalinflammation has been induced using Dextran Sulfate. Mice were treatedwith Vehicle (1% citric acid in water, 5 mL/kg) or the indicated dosesof compound E2 in μmol/kg.

FIG. 4 : Killing of phagocytosed Salmonella typhimurium by murinemacrophages treated with either the commercial antibiotic azithromycinor Compound E2. Compound E2 stimulates the killing of bacterial cells bymacrophages.

FIG. 5 : Effect of substance E5 on the response of mice to an infectionby Staphylococcus. Treatment with the substance E5 results in a fasterrecovery of weight due to faster clearance of bacteria

FIG. 6 : Effect of substances E2, E5 and Azithromycin on the ability ofmice to clear an infection by Staphylococcus aureus Newman. Bacteria arequantified as CFU recovered from a standard sample of kidney. Treatmentwith substances decreases recovered bacteria in a dose responsivemanner.

FIG. 7 : Effect of substances on the ability of mice to tolerate dextransulfate colitis. Cyclosporine is provided at a dose of 25 mg/kg, allother substances including azithromycin are provided at a dose of 0.1μmol/kg.

FIG. 8 : Effect of substances E5 and Cyclosporin on the liver weight ofmice treated with dextran sulfate colitis. Data are the mean of N=8 andare plotted with the 95% confidence interval.

FIG. 9 : Effect of substances E-5 and E-241 versus Vehicle on thedevelopment of EAE in the C57B6 mouse. Plotted is the clinical scorebased with antigen injected on day 0 and substance started on day 7.Data are the mean of N=8.

FIG. 10 : Effect of substances compared with the positive controlCyclosporin on the body weight of mice treated with dextran sulfatecolitis. Data are the mean of N=8 and are plotted with the 95%confidence interval. Above the bars are the p values vs. Vehicle for aT-test.

FIG. 11 : Effect of substances compared with the positive controlCyclosporin on the colon length of mice treated with dextran sulfatecolitis. Data are the mean of N=8 and are plotted with the 95%confidence interval. Above the bars are the p values vs. Vehicle for aT-test.

FIG. 12 : Effect of substances compared with the positive controlCyclosporin on the amount of fluorocein labelled dextran (FITC) taken upinto the serums of mice treated with dextran sulfate colitis. 4 h priorto sampling, mice are treated with an oral suspension of FITC dextranwhich would normally not enter the blood stream. The effect of DSS is todisrupt the gut epithelium allowing larger molecules to enter the bloodstream. Reductions in FITC dextran suggest improved barrier function.Data are the mean of N=8 and are plotted with the 95% confidenceinterval. Above the bars are the p values vs. Vehicle for a T-test.

FIG. 13 : Effect of substances compared with the positive controlCyclosporin on the serum Calcium of mice treated with dextran sulfate toinduce colitis at day 8 after starting DSS. Data are the mean of N=8 andare plotted with the 95% confidence interval. Above the bars are the pvalues vs. Vehicle for a T-test.

FIG. 14 : Effect of substances compared with the positive controlCyclosporin on the clinical score of mice treated with dextran sulfateto induce colitis at day 8 after starting DSS. Data are the mean of N=8and are plotted with the 95% confidence interval. Above the bars are thep values vs. Vehicle for a T-test.

FIG. 15 : Effect of substances compared with the positive controlCyclosporin on the serum Potassium of mice treated with dextran sulfateto induce colitis at day 8 after starting DSS. Data are the mean of N=8and are plotted with the 95% confidence interval. Above the bars are thep values vs. Vehicle for a T-test.

FIG. 16 : Effect of substances compared with the positive controlCyclosporin on the serum Total Bilirubin of mice treated with dextransulfate to induce colitis at day 8 after starting DSS. Data are the meanof N=8 and are plotted with the 95% confidence interval. Above the barsare the p values vs. Vehicle for a T-test.

FIG. 17 : Body weight of BALBc mice at day 7 after commencing 2.5% DSSin water. DSS causes lesions in the colon that lead to weight loss.Substance E-2, amongst others, protects against weight loss. Data arethe mean of N=8 and are plotted with the 95% confidence interval.

FIG. 18 : Body weight and clinical score of BALBc mice at day 9 aftercommencing 2.5% DSS in water. DSS causes lesions in the colon that leadto weight loss. Substances E-3, E-238 and E-553, amongst others,stimulate inflammation. All doses 1.34 μmol/kg. Data are the mean of N=8and are plotted with the 95% confidence interval. Above the bars are thep values vs. Vehicle for a T-test.

FIG. 19 : Body weight and clinical score of BALBc mice at day 7 aftercommencing 2.5% DSS in water. DSS causes lesions in the colon that leadto weight loss. Substance like E-51 with weight greater than Vehicleprotect against inflammation. Doses are as indicated. Data are the meanof N=8 and are plotted with the 95% confidence interval. Above the barsare the p values vs. Vehicle for a T-test.

FIG. 20 : The compounds containing R₁ nitrate ester have preferentialdistribution to the lung. Data show the concentration of the substancein the lung and liver at 6 h after administration of a 10 mg/kg dosep.o. in 2% citric acid.

FIG. 21 : The effect of various compounds on the rate of killing ofSalmonella typhimurium following incubation and phagocytosis by J774murine cells. The number of surviving bacteria is an indicator of thedegree of intracellular killing of the bacteria, All substances aresupplied at an initial concentration of 1 μM.

DESCRIPTION OF EMBODIMENTS

General Procedure for the Introduction of the Nitrooxide Group

The compound to be nitrated (1 equiv.) (—SH, —OH, —NH) is dissolved orsuspended in acetic acid (approximately 6.0 ml per 1 mmol compound to benitrated) and a solution of nitric acid (10% in acetic anhydride, about3.25 ml per 1 mmol compound to be nitrated) is slowly added to thesystem while cooling in an ice bath. When TLC indicated completeconsumption of starting materials the mixture is poured onto icehydrolyzing any remains of acetic anhydride, followed by cautiousneutralization of acid species with sodium bicarbonate. Extraction ofthe aqueous system with dichloromethane (3×), drying of combined organicphases over sodium sulfate and subsequent purification of crude productsby column chromatography (acetone-cyclohexane 1:3→1:1) yields productsas amorphous white foams.

The invention will be further described in the following intermediatesand examples. It should be understood that these examples are forillustrative purposes only and are not to be construed as limiting thisinvention in any, manner.

EXAMPLES

Unless otherwise specified, all commercially available reagents andsolvents were used without prior purification. All chemical structuresand names are generated from ChemDraw Ultra (Cambridge).

TABLE 1 Examples of ALC Core Entry ALC Core Structure A-1 Azithromycin(R₁ to R₅ = H, R₆ = CH₃)

A-2 Erythromycin (X = OH) (R₁ to R₅ = H, R₆ = CH₃) or ErythromycinN-Oxime (X = N—OH)

A-3 Hydroxychloroquine (R₁ = H, R₂ = CH₃)

A-4 N-ethanol HCQ (R₁ = H, R₂ = H)

A-5 Propranolol (R₁ = H, R₂ = CH₃)

A-6 N-ethanol-propranolol (R₁ = H, R₂ = H)

A-7 4-hydroxypropranolol (R₁ = H, R₂ = H, R₃ = H)

A-8 2-(4-fluorophenyl)-3-amino-4-(3-[2,3-di{butyroyloxy}propyloxy]phenyl)- carbonylpyrazole (R₁ = H, R₂ = H)

A-9 2-(4-pyridyl)-3-amino-4-(3-[2,3- di{butyroyloxy}propyloxy]-phenyl)carbonylpyrazole (R₁ = H, R₂ = H)

A-10 C5Y0073 (R₁ to R₅ = H, R₆ = CH₃) or14-(4-Dimethylamino-3-hydroxy-6-methyl-tetrahydro-pyran-2-yloxy)-5-ethyl-1,6,7-trihydroxy-2,6,8,9,11,13,15- heptamethyl-4,16-dioxa-9-aza-bicyclo[11.2.1]hexadecan-3-one

A-11/ E-16 CSY0041 (R₁ to R₆ = H) or2-Ethyl-3,4,10-trihydroxy-13-(5-hydroxy-4-methoxy-4,6-dimethyl-tetrahydro-pyran-2-yloxy)-11-(3-hydroxy-6-methyl-4-methylamino-tetrahydro-pyran-2-yloxy)-3,5,6,8,10,12,14-heptamethyl-1-oxa-6- aza-cyclopentadecan-15-one

A-12 C5Y1239 (R₁ to R₅ = H) or 11-(4-Dimethylamino-3-hydroxy-6-methyl-tetrahydro-pyran-2-yloxy)-2-ethyl- 3,4,10,13-tetrahydroxy-3,5,6,8,10,12,14-heptamethyl-1-oxa-6-aza- cyclopentadecan-15-one

A-13.1 CSY1130 (R₁ to R₆ = H) (preparation see Example or11-{4-[Bis-(2-hydroxy-ethyl)-amino]-3-hydroxy-6-methyl-tetrahydro-pyran-2-yloxy}-2-ethyl-3,4,10-trihydroxy-13-(5- hydroxy-4-methoxy-4,6-dimethyl-tetrahydro-pyran-2-yloxy)- 3,5,6,8,10,12,14-heptamethyl-1-oxa-6-aza-cyclopentadecan-15-one

A-13.2 CSY5632 (R₁ to R₆ = H) or (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-11-(((2S,3S,6R)-3-(bis(2- hydroxyethyl)amino)-4-hydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)-2- ethyl-3,4,10-trihydroxy-13-(((2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyltetrahydro-2H-pyran-2-yl)oxy)-3,5,6,8,10,12,14-heptamethyl-1-oxa-6- azacydopentadecan-15-one

A-14.1 CSY2219 (R₁ to R₅ = H) or2-Ethyl-3,4,10-trihydroxy-13-(5-hydroxy-4-methoxy-4,6-dimethyl-tetrahydro-pyran-2-yloxy)-11-(3-hydroxy-6-methyl-4-morpholin-4-yl-tetrahydro-pyran-2-yloxy)-3,5,6,8,10,12,14-heptamethyl-1- oxa-6-aza-cyclopentadecan-15-one

A-14.2 CSY5602 (R₁ to R₅ = H) or (2R,3S,4R,5R,8R,10R,11R,12S,13S,14R)-2-ethyl-3,4,10-trihydroxy-13- (((2R,4R,5S,6S)-5-hydroxy-4-methoxy-4,6-dimethyltetrahydro-2H-pyran-2-yl)oxy)-11-(((2S,3S,6R)-4-hydroxy-6-methyl-3- morpholinotetrahydro-2H-pyran-2-yl)oxy)-3,5,6,8,10,12,14-heptamethyl-1- oxa-6-azacyclopentadecan-15-one

A-15 CSY1019 (R₁ to R₅ = H) or 11-(4-Dimethylamino-6-methyl-3-nitrooxy-tetrahydro-pyran-2-yloxy)-2-ethyl-3,4,10-trihydroxy-13-(5-hydroxy-4-methoxy-4,6-dimethyl-tetrahydro-pyran-2-yloxy)-3,5,6,8,10,12,14-heptamethyl-1-oxa-6-aza-cyclopentadecan-15-one

A-16 Tildipirosin (R₁ to R₃ = H)

A-17 Gamithromycin (R₁ to R₅ = H)

A-18 Tylosin (R₁ to R₅ = H)

A-19.1 Polyamines (R₁ to R₂ = H)

A-19.2 Polyamines (R₁ to R₄ = H)

A-19.3 Polyamines (R₁ = R₂ = H; R₃ = alkyl, usually ethyl)

A-20.1 Tris(hydroxymethyl)nitromethane (R₁ to R₃ = H)

A-20.2 Sodium Tris(hydroxymethyl)aminopropylsulfonate (R₁ to R₃ = H)

A-21 Clarithromycin (R₁ to R₄ = H)

A-22 Tulathromycin (R₁ to R₅ = 0)

TABLE 2 Examples of compounds showing the appropriate substituents.Compound formulae based on “ALC core” structures as detailed in Table 1(above). Compound Entry ALC Core R₁ R₂ R₃ R₄ R₅ R₆ E-1 Azithromycin NO₂H H H H CH₃ E-2 Azithromycin NO₂ Ac H H H CH₃ E-3 Azithromycin NO₂ H H HH C-10 alkyl E-4 Azithromycin NO₂ Ac H H H C-10 alkyl E-5 AzithromycinNO₂ Propionyl H H H CH₃ E-6 Azithromycin p-Nitro NO₂ NO₂ H H CH₃ benzoylE-7 Azithromycin Benzoyl NO₂ H H H CH₃ E-8 Erythromycin NO₂ Ac H H H CH₃oxime E-9 Azithromycin Ac NO₂ H H H CH₃ E-10 Azithromycin H NO₂ H H HCH₃ E-11 Azithromycin NO₂ ch₃ H H H CH₃ E-12 Azithromycin H Tetranitro HH H CH₃ moiety¹ E-13 Azithromycin NO₂ Propionyl H H Propionyl CH₃ E-14Azithromycin H H H H H Propanol- NO₂ E-15 Azithromycin H H H H H CH₃E-16 Azithromycin H H H H H H E-17/I-7 Azithromycin H Ac H H H CH₃ E-18Azithromycin H Propionyl H H H CH₃ E-19 Azithromycin H Butyryl H H H CH₃E-20/I-2 Azithromycin H H H H H C-10 alkyl E-21 Azithromycin NO₂ NO₂ H HH CH₃ E-22/I-4 Azithromycin Ac CH₂CCH H H H CH₃ E-23/I-6 Azithromycin AcAc H H H CH₃ E-24 Azithromycin NO₂ Butyryl H H H CH₃ E-25//I-3Azithromycin Ac H H H H CH₃ E-26/I-8 Azithromycin Benzoyl H H H H CH₃E-27 Azithromycin Succinyl H H H H CH₃ E-28 Azithromycin Ac Propionyl HH H CH₃ E-29 Azithromycin Ac Butyryl H H H CH₃ E-30 AzithromycinPropionyl H H H H CH₃ E-31 Azithromycin Propionyl NO₂ H H H CH₃ E-32Azithromycin Propionyl Ac H H H CH₃ E-33 Azithromycin PropionylPropionyl H H H CH₃ E-34 Azithromycin Propionyl Butyryl H H H CH₃ E-35Azithromycin Butyryl H H H H CH₃ E-36 Azithromycin Butyryl NO₂ H H H CH₃E-37 Azithromycin Butyryl Ac H H H CH₃ E-38 Azithromycin ButyrylPropionyl H H H CH₃ E-39 Azithromycin Butyryl Butyryl H H H CH₃ E-40Azithromycin H Succinyl H H H CH₃ E-41 Azithromycin H Pyruvyl H H H CH₃E-42 Azithromycin H Maleyl H H H CH₃ E-43 Azithromycin H Lactyl H H HCH₃ E-44 Azithromycin H Isobutyryl H H H CH₃ E-45 Azithromycin H ValerylH H H CH₃ E-46 Azithromycin H Isovaleryl H H H CH₃ E-47 AzithromycinButyryl Butyryl Butyryl Butyryl H CH₃ E-48 Azithromycin Butyryl ButyrylButyryl H H CH₃ E-49 Azithromycin Ac Ac Ac Ac H CH₃ E-50 Azithromycin AcAc Ac H H CH₃ E-51 Azithromycin Propionyl Propionyl Propionyl PropionylH CH₃ E-52 Azithromycin Propionyl Propionyl Propionyl H H CH₃ E-53Azithromycin Succinyl Succinyl Succinyl Succinyl H CH₃ E-54 AzithromycinPyruvyl Pyruvyl Pyruvyl Pyruvyl H CH₃ E-55 Azithromycin Maleyl MaleylMaleyl Maleyl H CH₃ E-56 Azithromycin Lactyl Lactyl Lactyl Lactyl H CH₃E-57 Azithromycin Isobutyryl Isobutyryl Isobutyryl Isobutyryl H CH₃ E-58Azithromycin Valeryl Valeryl Valeryl Valeryl H CH₃ E-59 AzithromycinIsovaleryl Isovaleryl Isovaleryl Isovaleryl H CH₃ E-60 AzithromycinSuccinyl Succinyl Succinyl H H CH₃ E-61 Azithromycin Pyruvyl PyruvylPyruvyl H H CH₃ E-62 Azithromycin Maleyl Maleyl Maleyl H H CH₃ E-63Azithromycin Lactyl Lactyl Lactyl H H CH₃ E-64 Azithromycin IsobutyrylIsobutyryl Isobutyryl H H CH₃ E-65 Azithromycin Valeryl Valeryl ValerylH H CH₃ E-66 Azithromycin Isovaleryl Isovaleryl Isovaleryl H H CH₃ E-67Azithromycin Succinyl Succinyl H H H CH₃ E-68 Azithromycin PyruvylPyruvyl H H H CH₃ E-69 Azithromycin Maleyl Maleyl H H H CH₃ E-70Azithromycin Lactyl Lactyl H H H CH₃ E-71 Azithromycin IsobutyrylIsobutyryl H H H CH₃ E-72 Azithromycin Valeryl Valeryl H H H CH₃ E-73Azithromycin Isovaleryl Isovaleryl H H H CH₃ E-74 AzithromycinAcetoxypropionyl H H H H CH₃ E-75 Azithromycin Lipoyl Lipoyl H H H CH₃E-76 Azithromycin H Lipoyl H H H CH₃ E-77 Azithromycin Lipoyl H H H HCH₃ E-78 Azithromycin NO₂ Lipoyl H H H CH₃ E-79 Azithromycin NO₂Succinyl - H H H CH₃ dithiole- 3-thione E-80 Azithromycin Succinyl - H HH H CH₃ dithiole- 3-thione E-81 Azithromycin Polysulfide H H H H CH₃ethyl carbonate E-82 Azithromycin NO- H H H H CH₃ thioethylcarbonateE-83 Azithromycin O-Phenyl H H H H CH₃ chlorothionocarbonate E-84Azithromycin n- H H H H CH₃ hexanoyl E-85 AzithromycinBromoethylcarbonate H H H H CH₃ E-86 Azithromycin Vinyll H H H H CH₃carbonate E-87 Hydroxychloroquine Ac H E-88 Hydroxychloroquine PropionylH E-89 Hydroxychloroquine Butyryl H E-90 Hydroxychloroquine Succinyl HE-91 Hydroxychloroquine Pyruvyl H E-92 Hydroxychloroquine Maleyl H E-93Hydroxychloroquine Lactyl H E-94 Hydroxychloroquine Isobutyryl H E-95Hydroxychloroquine Valeryl H E-96 Hydroxychloroquine Isovaleryl H E-97Hydroxychloroquine Lipoyl H E-98 Hydroxychloroquine Ac CH₃ E-99Hydroxychloroquine Propionyl CH₃ E-100 Hydroxychloroquine Butyryl CH₃E-101 Hydroxychloroquine Succinyl CH₃ E-102 Hydroxychloroquine PyruvylCH₃ E-103 Hydroxychloroquine Maleyl CH₃ E-104 Hydroxychloroquine LactylCH₃ E-105 Hydroxychloroquine Isobutyryl CH₃ E-106 HydroxychloroquineValeryl CH₃ E-107 Hydroxychloroquine Isovaleryl CH₃ E-108Hydroxychloroquine Lipoyl CH₃ E-109 N-ethanol HCQ NO₂ Ac E-110 N-ethanolHCQ NO₂ Propionyl E-111 N-ethanol HCQ NO₂ Butyryl E-112 N-ethanol HCQNO₂ Succinyl E-113 N-ethanol HCQ NO₂ Pyruvyl E-114 N-ethanol HCQ NO₂Maleyl E-115 N-ethanol HCQ NO₂ Lactyl E-116 N-ethanol HCQ NO₂ IsobutyrylE-117 N-ethanol HCQ NO₂ Valeryl E-118 N-ethanol HCQ NO₂ Isovaleryl E-119N-ethanol HCQ NO₂ Lipoyl E-120 N-ethanol HCQ Ac NO₂ E-121 N-ethanol HCQPropionyl NO₂ E-122 N-ethanol HCQ Butyryl NO₂ E-123 N-ethanol HCQSuccinyl NO₂ E-124 N-ethanol HCQ Pyruvyl NO₂ E-125 N-ethanol HCQ MaleylNO₂ E-126 N-ethanol HCQ Lactyl NO₂ E-127 N-ethanol HCQ Isobutyryl NO₂E-128 N-ethanol HCQ Valeryl NO₂ E-129 N-ethanol HCQ Isovaleryl NO₂ E-130N-ethanol HCQ Lipoyl NO₂ E-131 Propranolol Ac H E-132 PropranololPropionyl H E-133 Propranolol Butyryl H E-134 Propranolol Succinyl HE-135 Propranolol Pyruvyl H E-136 Propranolol Maleyl H E-137 PropranololLactyl H E-138 Propranolol Isobutyryl H E-139 Propranolol Valeryl HE-140 Propranolol Isovaleryl H E-141 Propranolol Lipoyl H E-142Propranolol 2-O- H Nitrolactyl E-143 Propranolol Ac CH₃ E-144Propranolol Propionyl CH₃ E-145 Propranolol Butyryl CH₃ E-146Propranolol Succinyl CH₃ E-147 Propranolol Pyruvyl CH₃ E-148 PropranololMaleyl CH₃ E-149 Propranolol Lactyl CH₃ E-150 Propranolol Isobutyryl CH₃E-151 Propranolol Valeryl CH₃ E-152 Propranolol Isovaleryl CH₃ E-153Propranolol Lipoyl CH₃ E-154 N-ethanol- NO₂ Ac propranolol E-155N-ethanol- NO₂ Propionyl propranolol E-156 N-ethanol- NO₂ Butyrylpropranolol E-157 N-ethanol- NO₂ Succinyl propranolol E-158 N-ethanol-NO₂ Pyruvyl propranolol E-159 N-ethanol- NO₂ Maleyl propranolol E-160N-ethanol- NO₂ Lactyl propranolol E-161 N-ethanol- NO₂ Isobutyrylpropranolol E-162 N-ethanol- NO₂ Valeryl propranolol E-163 N-ethanol-NO₂ Isovaleryl propranolol E-164 N-ethanol- NO₂ Lipoyl propranolol E-165N-ethanol- Propionyl NO₂ propranolol E-166 N-ethanol- Butyryl NO₂propranolol E-167 N-ethanol- Succinyl NO₂ propranolol E-168 N-ethanol-Pyruvyl NO₂ propranolol E-169 N-ethanol- Maleyl NO₂ propranolol E-170N-ethanol- Lactyl NO₂ propranolol E-171 N-ethanol- Isobutyryl NO₂propranolol E-172 N-ethanol- Valeryl NO₂ propranolol E-173 N-ethanol-Isovaleryl NO₂ propranolol E-174 N-ethanol- Lipoyl NO₂ propranolol E-1754-hydroxy NO₂ H Ac propranolol E-176 4-hydroxy NO₂ H Propionylpropranolol E-177 4-hydroxy NO₂ H Butyryl propranolol E-178 4-hydroxyNO₂ H Succinyl propranolol E-179 4-hydroxy NO₂ H Pyruvyl propranololE-180 4-hydroxy NO₂ H Maleyl propranolol E-181 4-hydroxy NO₂ H Lactylpropranolol E-182 4-hydroxy NO₂ H Isobutyryl propranolol E-183 4-hydroxyNO₂ H Valeryl propranolol E-184 4-hydroxy NO₂ H Isovaleryl propranololE-185 4-hydroxy NO₂ H Lipoyl propranolol E-186 4-hydroxy NO₂ CH₃ Acpropranolol E-187 4-hydroxy NO₂ CH₃ Propionyl propranolol E-1884-hydroxy NO₂ CH₃ Butyryl propranolol E-189 4-hydroxy NO₂ CH₃ Succinylpropranolol E-190 4-hydroxy NO₂ CH₃ Pyruvyl propranolol E-191 4-hydroxyNO₂ CH₃ Maleyl propranolol E-192 4-hydroxy NO₂ CH₃ Lactyl propranololE-193 4-hydroxy NO₂ CH₃ Isobutyryl propranolol E-194 4-hydroxy NO₂ CH₃Valeryl propranolol E-195 4-hydroxy NO₂ CH₃ Isovaleryl propranolol E-1964-hydroxy NO₂ CH₃ Lipoyl propranolol E-197 A-8 Ac Ac E-198 A-8 PropionylPropionyl E-199 A-8 Butyryl Butyryl E-200 A-8 Succinyl Succinyl E-201A-8 Pyruvyl Pyruvyl E-202 A-8 Maleyl Maleyl E-203 A-8 Lactyl LactylE-204 A-8 Isobutyryl Isobutyryl E-205 A-8 Valeryl Valeryl E-206 A-8Isovaleryl Isovaleryl E-207 A-8 Lipoyl Lipoyl E-208 A-9 Ac Ac E-209 A-9Propionyl Propionyl E-210 A-9 Butyryl Butyryl E-211 A-9 SuccinylSuccinyl E-212 A-9 Pyruvyl Pyruvyl E-213 A-9 Maleyl Maleyl E-214 A-9Lactyl Lactyl E-215 A-9 Isobutyryl Isobutyryl E-216 A-9 Valeryl ValerylE-217 A-9 Isovaleryl Isovaleryl E-218 A-9 Lipoyl Lipoyl E-219 A-10 HTetranitro H H CH₃ moiety1 E-220 A-10 H H H H Propanol- NO₂ E-221 A-10 HH H H CH₃ E-222 A-10 H H H H H E-223 A-10 H Ac H H CH₃ E-224 A-10 HPropionyl H H CH₃ E-225 A-10 H Butyryl H H CH₃ E-226 A-10 H H H H C-10alkyl E-227 A-10 Ac CH₂CCH H H CH₃ E-228 A-10 Ac Ac H H CH₃ E-229 A-10Ac H H H CH₃ E-230 A-10 Benzoyl H H H CH₃ E-231 A-10 Succinyl H H H CH₃E-232 A-10 Ac Propionyl H H CH₃ E-233 A-10 Ac Butyryl H H CH₃ E-234 A-10Propionyl H H H CH₃ E-235 A-10 Propionyl Ac H H CH₃ E-236 A-10 PropionylPropionyl H H CH₃ E-237 A-10 Propionyl Butyryl H H CH₃ E-238 A-10Butyryl H H H CH₃ E-239 A-10 Butyryl Ac H H CH₃ E-240 A-10 ButyrylPropionyl H H CH₃ E-241 A-10 Butyryl Butyryl H H CH₃ E-242 A-10 HSuccinyl H H CH₃ E-243 A-10 H Pyruvyl H H CH₃ E-244 A-10 H Maleyl H HCH₃ E-245 A-10 H Lactyl H H CH₃ E-246 A-10 H Isobutyryl H H CH₃ E-247A-10 H Valeryl H H CH₃ E-248 A-10 H Isovaleryl H H CH₃ E-249 A-10Butyryl Butyryl Butyryl H CH₃ E-250 A-10 Butyryl Butyryl H H CH₃ E-251A-10 Ac Ac Ac H CH₃ E-252 A-10 Ac Ac H H CH₃ E-253 A-10 PropionylPropionyl Propionyl H CH₃ E-254 A-10 Propionyl Propionyl H H CH₃ E-255A-11/E-16 Butyric H H H H Butyric E-256 A-11/E-16 Butyric Butyric H H HButyric E-257 A-11/E-16 Butyric Butyric Butyric H H Butyric E-258A-11/E-16 H H H H H Mannose E-259 A-11/E-16 Propionic H H H H PropionicE-260 A-11/E-16 Propionic Propionic H H H Propionic E-261 A-11/E-16Propionic Propionic Propionic H H Propionic E-262 A-11/E-16 Ac H H H HAc E-263 A-11/E-16 Ac Ac H H H Ac E-264 A-11/E-16 Ac Ac Ac H H Ac E-265A-12 NO2 H H H H E-266 A-12 Ac Ac H H H E-267 A-12 Ac Ac Ac H H E-268A-12 Ac Ac Ac Ac H E-269 A-12 Propionic Propionic H H H E-270 A-12Propionic Propionic Propionic H H E-271 A-12 Propionic PropionicPropionic Propionic H E-272 A-12 Butyric Butyric H H H E-273 A-12Butyric Butyric Butyric H H E-274 A-12 Butyric Butyric Butyric Butyric HE-275 A-12 Succinic Succinic H H H E-276 A-12 Pyruvic Pyruvic H H HE-277 A-12 Maleic Maleic H H H E-278 A-12 Lactic Lactic H H H E-279 A-12Isobutyric Isobutyric H H H E-280 A-12 Isobutyric Isobutyric IsobutyricH H E-281 A-12 Isobutyric Isobutyric Isobutyric Isobutyric H E-282 A-12Valeric Valeric H H H E-283 A-12 Valeric Valeric Valeric H H E-284 A-12Valeric Valeric Valeric Valeric H E-285 A-12 Isovaleric Isovaleric H H HE-286 A-12 Isovaleric Isovaleric Isovaleric H H E-287 A-12 IsovalericIsovaleric Isovaleric Isovaleric H E-288 A-12 Lipoic Lipoic H H H E-289A-12 Lipoic Lipoic Lipoic H H E-290 A-12 Lipoic Lipoic Lipoic Lipoic HE-291 A-13.1 H H H H H Ac E-292 A-13.1 Ac H H H H Ac E-293 A-13.1 Ac AcH H H Ac E-294 A-13.1 Ac Ac Ac H H Ac E-295 A-13.1 H H H H H PropionicE-296 A-13.1 Propionic H H H H Propionic E-297 A-13.1 PropionicPropionic H H H Propionic E-298 A-13.1 Propionic Propionic Propionic H HPropionic E-299 A-13.1 H H H H H Butyric E-300 A-13.1 Butyric H H H HButyric E-301 A-13.1 Butyric Butyric H H H Butyric E-302 A-13.1 ButyricButyric Butyric H H Butyric E-303 A-13.1 H H H H H Succinic E-304 A-13.1H H H H H Pyruvic E-305 A-13.1 H H H H H Maleic E-306 A-13.1 H H H H HLactic E-307 A-13.1 H H H H H Isobutyric E-308 A-13.1 Isobutyric H H H HIsobutyric E-309 A-13.1 Isobutyric Isobutyric H H H Isobutyric E-310A-13.1 Isobutyric Isobutyric Isobutyric H H Isobutyric E-311 A-13.1 H HH H H Valeric E-312 A-13.1 Valeric H H H H Valeric E-313 A-13.1 ValericValeric H H H Valeric E-314 A-13.1 Valeric Valeric Valeric H H ValericE-315 A-13.1 H H H H H Isovaleric E-316 A-13.1 Isovaleric H H H HIsovaleric E-317 A-13.1 Isovaleric Isovaleric H H H Isovaleric E-318A-13.1 Isovaleric Isovaleric Isovaleric H H Isovaleric E-319 A-13.1 H HH H H Lipoic E-320 A-13.1 Lipoic H H H H Lipoic E-321 A-13.1 LipoicLipoic H H H Lipoic E-322 A-13.1 Lipoic Lipoic Lipoic H H Lipoic E-323A-13.2 H H H H H Ac E-324 A-13.2 Ac H H H H Ac E-325 A-13.2 Ac Ac H H HAc E-326 A-13.2 Ac Ac Ac H H Ac E-327 A-13.2 H H H H H Propionic E-328A-13.2 Propionic H H H H Propionic E-329 A-13.2 Propionic Propionic H HH Propionic E-330 A-13.2 Propionic Propionic Propionic H H PropionicE-331 A-13.2 H H H H H Butyric E-332 A-13.2 Butyric H H H H ButyricE-333 A-13.2 Butyric Butyric H H H Butyric E-334 A-13.2 Butyric ButyricButyric H H Butyric E-335 A-13.2 H H H H H Succinic E-336 A-13.2 H H H HH Pyruvic E-337 A-13.2 H H H H H Maleic E-338 A-13.2 H H H H H LacticE-339 A-13.2 H H H H H Isobutyric E-340 A-13.2 Isobutyric H H H HIsobutyric E-341 A-13.2 Isobutyric Isobutyric H H H Isobutyric E-342A-13.2 Isobutyric Isobutyric Isobutyric H H Isobutyric E-343 A-13.2 H HH H H Valeric E-344 A-13.2 Valeric H H H H Valeric E-345 A-13.2 ValericValeric H H H Valeric E-346 A-13.2 Valeric Valeric Valeric H H ValericE-347 A-13.2 H H H H H Isovaleric E-348 A-13.2 Isovaleric H H H HIsovaleric E-349 A-13.2 Isovaleric Isovaleric H H H Isovaleric E-350A-13.2 Isovaleric Isovaleric Isovaleric H H Isovaleric E-351 A-13.2 H HH H H Lipoic E-352 A-13.2 Lipoic H H H H Lipoic E-353 A-13.2 LipoicLipoic H H H Lipoic E-354 A-13.2 Lipoic Lipoic Lipoic H H Lipoic E-355A-14.1 Ac H H H H E-356 A-14.1 Ac Ac H H H E-357 A-14.1 Ac Ac Ac H HE-358 A-14.1 Propionic H H H H E-359 A-14.1 Propionic Propionic H H HE-360 A-14.1 Propionic Propionic Propionic H H E-361 A-14.1 Butyric H HH H E-362 A-14.1 Butyric Butyric H H H E-363 A-14.1 Butyric ButyricButyric H H E-364 A-14.1 Succinic H H H H E-365 A-14.1 Pyruvic H H H HE-366 A-14.1 Maleic H H H H E-367 A-14.1 Lactic H H H H E-368 A-14.1Isobutyric H H H H E-369 A-14.1 Isobutyric Isobutyric H H H E-370 A-14.1Isobutyric Isobutyric Isobutyric H H E-371 A-14.1 Valeric H H H H E-372A-14.1 Valeric Valeric H H H E-373 A-14.1 Valeric Valeric Valeric H HE-374 A-14.1 Isovaleric H H H H E-375 A-14.1 Isovaleric Isovaleric H H HE-376 A-14.1 Isovaleric Isovaleric Isovaleric H H E-377 A-14.1 Lipoic HH H H E-378 A-14.1 Lipoic Lipoic H H H E-379 A-14.1 Lipoic Lipoic LipoicH H E-380 A-14.2 Ac H H H H E-381 A-14.2 Ac Ac H H H E-382 A-14.2 Ac AcAc H H E-383 A-14.2 Propionic H H H H E-384 A-14.2 Propionic Propionic HH H E-385 A-14.2 Propionic Propionic Propionic H H E-386 A-14.2 ButyricH H H H E-387 A-14.2 Butyric Butyric H H H E-388 A-14.2 Butyric ButyricButyric H H E-389 A-14.2 Succinic H H H H E-390 A-14.2 Pyruvic H H H HE-391 A-14.2 Maleic H H H H E-392 A-14.2 Lactic H H H H E-393 A-14.2Isobutyric H H H H E-394 A-14.2 Isobutyric Isobutyric H H H E-395 A-14.2Isobutyric Isobutyric Isobutyric H H E-396 A-14.2 Valeric H H H H E-397A-14.2 Valeric Valeric H H H E-398 A-14.2 Valeric Valeric Valeric H HE-399 A-14.2 Isovaleric H H H H E-400 A-14.2 Isovaleric Isovaleric H H HE-401 A-14.2 Isovaleric Isovaleric Isovaleric H H E-402 A-14.2 Lipoic HH H H E-403 A-14.2 Lipoic Lipoic H H H E-404 A-14.2 Lipoic Lipoic LipoicH H E-405 A-15 Succinic H H H E-406 A-15 Pyruvic H H H E-407 A-15 MaleicH H H E-408 A-15 Lactic H H H E-409 A-15 Isobutyric H H H E-410 A-15Isobutyric Isobutyric H H E-411 A-15 Valeric H H H E-412 A-15 ValericValeric H H E-413 A-15 Isovaleric H H H E-414 A-15 Isovaleric IsovalericH H E-415 A-15 Lipoic Lipoic H H E-416 A-16 Ac H H E-417 A-16 Ac Ac HE-418 A-16 Ac Ac Ac E-419 A-16 Propionic H H E-420 A-16 PropionicPropionic H E-421 A-16 Propionic Propionic Propionic E-422 A-16 ButyricH H E-423 A-16 Butyric Butyric H E-424 A-16 Butyric Butyric ButyricE-425 A-16 Isobutyric H H E-426 A-16 Isobutyric Isobutyric H E-427 A-16Isobutyric Isobutyric Isobutyric E-428 A-16 Valeric H H E-429 A-16Valeric Valeric H E-430 A-16 Valeric Valeric Valeric E-431 A-16Isovaleric H H E-432 A-16 Isovaleric Isovaleric H E-433 A-16 IsovalericIsovaleric Isovaleric E-434 A-16 Lipoic H H E-435 A-16 Lipoic Lipoic HE-436 A-16 Lipoic Lipoic Lipoic E-437 A-16 Hexanoic H H E-438 A-16Hexanoic Hexanoic H E-439 A-16 Hexanoic Hexanoic Hexanoic E-440 A-16Heptanoic H H E-441 A-16 Heptanoic Heptanoic H E-442 A-16 HeptanoicHeptanoic Heptanoic E-443 A-16 Octanoic H H E-444 A-16 Octanoic OctanoicH E-445 A-16 Octanoic Octanoic Octanoic E-446 A-16 Decanoic H H E-447A-16 Decanoic Decanoic H E-448 A-16 Decanoic Decanoic Decanoic E-449A-16 Dodecanoic H H E-450 A-16 Dodecanoic Dodecanoic H E-451 A-16Dodecanoic Dodecanoic Dodecanoic E-452 A-17 Ac H H H H E-453 A-17 Ac AcH H H E-454 A-17 Ac Ac Ac H H E-455 A-17 Propionic H H H H E-456 A-17Propionic Propionic H H H E-457 A-17 Propionic Propionic Propionic H HE-458 A-17 Butyric H H H H E-459 A-17 Butyric Butyric H H H E-460 A-17Butyric Butyric Butyric H H E-461 A-17 Isobutyric H H H H E-462 A-17Isobutyric Isobutyric H H H E-463 A-17 Isobutyric Isobutyric IsobutyricH H E-464 A-17 Valeric H H H H E-465 A-17 Valeric Valeric H H H E-466A-17 Valeric Valeric Valeric H H E-467 A-17 Isovaleric H H H H E-468A-17 Isovaleric Isovaleric H H H E-469 A-17 Isovaleric IsovalericIsovaleric H H E-470 A-17 Lipoic H H H H E-471 A-17 Lipoic Lipoic H H HE-472 A-17 Lipoic Lipoic Lipoic H H E-473 A-17 Hexanoic H H H H E-474A-17 Hexanoic Hexanoic H H H E-475 A-17 Hexanoic Hexanoic Hexanoic H HE-476 A-17 Heptanoic H H H H E-477 A-17 Heptanoic Heptanoic H H H E-478A-17 Heptanoic Heptanoic Heptanoic H H E-479 A-17 Octanoic H H H H E-480A-17 Octanoic Octanoic H H H E-481 A-17 Octanoic Octanoic Octanoic H HE-482 A-17 Decanoic H H H H E-483 A-17 Decanoic Decanoic H H H E-484A-17 Decanoic Decanoic Decanoic H H E-485 A-17 Dodecanoic H H H H E-486A-17 Dodecanoic Dodecanoic H H H E-487 A-17 Dodecanoic DodecanoicDodecanoic H H E-488 A-18 Ac H H H H E-489 A-18 Ac Ac H H H E-490 A-18Ac Ac Ac H H E-491 A-18 Propionic H H H H E-492 A-18 Propionic PropionicH H H E-493 A-18 Propionic Propionic Propionic H H E-494 A-18 Butyric HH H H E-495 A-18 Butyric Butyric H H H E-496 A-18 Butyric ButyricButyric H H E-497 A-18 Isobutyric H H H H E-498 A-18 IsobutyricIsobutyric H H H E-499 A-18 Isobutyric Isobutyric Isobutyric H H E-500A-18 Valeric H H H H E-501 A-18 Valeric Valeric H H H E-502 A-18 ValericValeric Valeric H H E-503 A-18 Isovaleric H H H H E-504 A-18 IsovalericIsovaleric H H H E-505 A-18 Isovaleric Isovaleric Isovaleric H H E-506A-18 Lipoic H H H H E-507 A-18 Lipoic Lipoic H H H E-508 A-18 LipoicLipoic Lipoic H H E-509 A-18 Hexanoic H H H H E-510 A-18 HexanoicHexanoic H H H E-511 A-18 Hexanoic Hexanoic Hexanoic H H E-512 A-18Heptanoic H H H H E-513 A-18 Heptanoic Heptanoic H H H E-514 A-18Heptanoic Heptanoic Heptanoic H H E-515 A-18 Octanoic H H H H E-516 A-18Octanoic Octanoic H H H E-517 A-18 Octanoic Octanoic Octanoic H H E-518A-18 Decanoic H H H H E-519 A-18 Decanoic Decanoic H H H E-520 A-18Decanoic Decanoic Decanoic H H E-521 A-18 Dodecanoic H H H H E-522 A-18Dodecanoic Dodecanoic H H H E-523 A-18 Dodecanoic Dodecanoic DodecanoicH H E-524 A-19.1 Ac Ac E-525 A-19.1 Propionic Propionic E-526 A-19.1Butyric Butyric E-527 A-19.1 Isobutyric Isobutyric E-528 A-19.1 ValericValeric E-529 A-19.1 Isovaleric Isovaleric E-530 A-19.1Adamantylcarboxyl Adamantylcarboxyl E-531 A-19.2 Ac Ac Ac Ac E-532A-19.2 Propionic Propionic Propionic Propionic E-533 A-19.2 ButyricButyric Butyric Butyric E-534 A-19.2 Isobutyric Isobutyric IsobutyricIsobutyric E-535 A-19.2 Valeric Valeric Valeric Valeric E-536 A-19.2Isovaleric Isovaleric Isovaleric Isovaleric E-537 A-20.1 Butyric ButyricButyric E-538 A-20.1 Ac Ac Ac E-539 A-20.1 Propionic Propionic PropionicE-540 A-1 N-Phenyl H H H H CH3 chlorothionoformate E-541 A-1 Imiquimod-H H H H CH3 Succinate E-542 A-1 Resiquimod- H H H H CH3 Succinate E-543A-1 Succinate- H H H H CH3 ethyl ester E-544 A-1 Indole-3- H H H H CH3propionic E-545 A-1 Cyclopropanecarboxylic H H H H CH3 E-546 A-1Cyclobutanecarboxylic H H H H CH3 E-547 A-1 Nicotinic H H H H CH3 E-548A-1 Chenodeoxycholic H H H H CH3 E-549 A-1 Ferrocenylacetic H H H H CH3E-550 A-1 Lipoic-S H H H H CH3 derivatives E-551 A-1 Methoxyacetic H H HH CH3 E-552 C2 (see Table 16) Butyric Butyric E-553 A-1 H ButyricButyric H H CH3 E-554 A-1 H Ac Ac H H CH3 E-555 A-1 H PropionicPropionic H H CH3 E-556 A-1 O-Acetyl H H H CH3 Lactic E-557 A-2 (X = O)Isovaleric H H H H CH3 E-558 A-2 (X = O) Valeric H H H H CH3 E-559 A-1Methoxyacetic Methoxyacetic Methoxyacetic Methoxyacetic H CH3 E-560 A-1Cyclobutanecarboxylic Cyclobutanecarboxylic H H H CH3 E-561 A-1Cyclobutanecarboxylic Cyclobutanecarboxylic Cyclobutanecarboxylic H HCH3 E-562 A-1 Nicotinic Nicotinic H H H CH3 E-563 A-1 NicotinicNicotinic Nicotinic H H CH3 E-564 A-2 (X = O) Ac H H H H CH3 E-566 A-10Isobutyric Isobutyric Isobutyric H CH3 E-567 A-10 Isobutyric IsobutyricH H CH3 E-568 A-10 Isobutyric H H H CH3 E-569 A-10 Valeric ValericValeric H CH3 E-570 A-10 Valeric Valeric H H CH3 E-571 A-10 Valeric H HH CH3 E-572 A-10 Isovaleric Isovaleric Isovaleric H CH3 E-573 A-10Isovaleric Isovaleric H H CH3 E-574 A-10 Isovaleric H H H CH3 E-575A-11/E-16 Butyric H H H Butyric Butyric E-576 A-19.1 Ac H E-577 A-19.1Propionic H E-578 A-19.1 Butyric H E-579 A-20.2 Butyric Butyric ButyricE-580 A-20.2 Ac Ac Ac E-581 A-20.2 Propionic Propionic Propionic E-582A-12 Valeric H H H H E-583 A-12 Isovaleric H H H H E-584 A-19.3 ValericH H E-585 A-19.3 Valeric H Ethyl E-586 A-19.3 Valeric Valeric H E-587A-19.3 Valeric Valeric Ethyl E-588 A-19.3 Isovaleric H H E-589 A-19.3Isovaleric H Ethyl E-590 A-19.3 Isovaleric Isovaleric H E-591 A-19.3Isovaleric Isovaleric Ethyl E-592 A-19.3 Butyric H H E-593 A-19.3Butyric H Ethyl E-594 A-19.3 Butyric Butyric H E-595 A-19.3 ButyricButyric Ethyl E-596 A-17 NO₂ H H H H E-597 A-17 NO₂ NO₂ H H H E-598 A-18H H NO₂ H H E-599 A-18 H NO₂ NO₂ H H E-600 A-11/E-16 H H no cladinose HH Mannose ring E-601 A-21 NO₂ E-602 A-21 NO₂ NO₂ E-603 A-21 NO₂ NO₂ HNO₂

Procedures

Example 1

Synthesis of E-1: Typical Nitration Procedure

Method 1. Acetic acid (40 mL) and Azithromycin (5 g, 6.73 mmol) werecharged in a round bottom flask. Initially, the reaction solidifiedwhich eventually during stifling, produced a homogenous solution. Theresulting solution was cooled in an ice-bath. Acetic anhydride (19.8 mL,209.5 mmol) was taken up in another reaction flask and cooled in an inice bath. To this was added dropwise, nitric acid (2.2 mL, 46.43 mmol).After complete addition, the mixture was transferred to a droppingfunnel and attached to the first reaction vessel containing themacrolide. The HNO₃-Ac₂O mixture was slowly added to the reaction (ca. 1drop per second). After complete addition, the reaction was allowed towarm to room temperature where it was stirred until reaction completion(3 h). The reaction was poured onto a stirred 200 mL ice-water. Stirringwas continued until the ice is completely melted. The resulting aqueoussolution was neutralized at first with a saturated solution of NaHCO₃,followed by pure solid NaHCO₃ to pH 8 to 9. The aqueous solution wasextracted with DCM (5×). The DCM extracts were dried (Na₂SO₄),evaporated in vacuo. The crude product was purified by columnchromatography (3:1 cyclohexane, ethyl acetate, 1% triethylamine) to getcompound E-1 as a white foam (30% yield).

These nitration conditions can be applied to other ALCs, and in caseswhere there is more than one reactive hydroxy species, selectiveprotection is necessary.

Method 2. Hydroxyalkyl species (1 mmol) was suspended in acetonitrile ina round bottom flask while stirring (magnetic stir bar, 300 rpm). Silvernitrate (2 eq. per hydroxyl group) was added and the mixture was cooledin an ice bath. Phosgene (solution in toluene, 1 eq. per hydroxyl group)was carefully added dropwise. Immediate precipitation of silver chlorideand formation of carbon dioxide indicated formation of nitro donor(caution: too quick CO₂ formation may result in strong foaming. Do notseal the flask!). After a couple of minutes a yellow color was obtainedand stirring was continued for 15 minutes. When ESI-MS indicatedsatisfying turn-over rate of starting materials the reaction wasquenched by addition of methanol, converting excess nitro donor tovolatile methyl nitrate. The system was diluted by addition of DCM andwas subject to extraction with saturated sodium bicarbonate solution(3×). Separation of organic phase, drying over sodium sulfate andevaporation of any volatiles in vacuo yielded the product as colorlessoil or beige to off-white foam.

TABLE 3 Nitration Examples Compound Synthesis Degree of Entry Method ALCSubstitution Yield MS E-596 2 A-17 Mono nitro n.d.* 822, M + H⁺ E-597 2A-17 Di nitro n.d.* 867, M + H⁺ 2 A-18 Oxidation n.d.* 954, M + Na⁺E-598 2 A-18 Mono nitro n.d.* 999, M + Na⁺ E-599 2 A-18 Di nitro n.d.*1045, M + Na⁺ E-601 2 A-21 Mono nitro n.d.* 793, M + H⁺ E-602 2 A-21 Dinitro n.d.* 838, M + H⁺ E-603 2 A-21 Tri nitro n.d.* 883, M + H⁺ n.d.*not determined

Example 2

Synthesis of E-2

Compound E-1 (791 mg, 1.0 mmol) was taken up in 15 mL dichloromethane.Pyridine (89 mL, 1.1 mmol) was added and the resulting solution wascooled in an ice bath for approximately 10 minutes. At this point, asolution of acetic anhydride (113 ml, 1.2 mmol) in dichloromethane (15mL) was added dropwise. The reaction was stirred continually at thistemperature and then progressively warmed to room temperature where itwas stirred overnight. The reaction was washed with a saturated solutionof ammonium chloride (3×), water (3×) and dried over anhydrous Na₂SO₄.The solvent was evaporated in vacuo. Co-evaporation with toluene isnecessary to remove residual pyridine from the system. This was followedby re-dissolving the residue in DCM and solvent evaporation twice toproduce a white foam, which was dried under high-vacuum to produce E-2(631 mg, 76%).

Example 3

Synthesis of E-3

E-3 was synthesized using standard nitration procedure described abovestarting from E-20.

Example 4

Synthesis of E-4

A solution of E-3 (1 mmol) and pyridine (1 mmol) in DCM (10 ml) wastreated with acetic anhydride (1.5 mmol) at ambient temperature.Stirring was continued until TLC (acetone-cyclohexane 1:3) indicatedcomplete consumption of the starting materials. The system was extractedwith an aqueous solution of NH4Cl (2× 10 ml) and water (3× 10 ml). Afterdrying over Na₂SO₄ all volatile components were evaporated in vacuo,traces of pyridine species were removed by co-evaporation with toluene(2×). The product E-4 was a colorless oil (41%).

Example 5

Synthesis of E-5

Method 1. A solution of propionic acid (1.2 mmol) in 1.2-dichloroethanewas treated with 1-Ethyl-3-(3-dimethyl-aminopropyl)carbodiimid (1.2mmol) in the presence of a catalytical amount of DMAP for 30 min. atambient temperature. Temperature was raised to 55°C., E-1 (1 mmol) wasadded and stirring was continued until TLC (acetone-cyclohexane 1:3)indicated complete consumption of the starting materials. The system wasextracted with water (3× 10 ml). After drying over Na₂SO₄ all volatilecomponents were evaporated in vacuo, traces of pyridine species wereremoved by co-evaporation with toluene (2×). The crude products werepurified by column chromatography (acetone-cyclohexane 1:3), yieldingthe product E-5 as white amorphous foam (39%).

Alternative Synthesis: Compound E-1 (310 mg, 0.39 mmol) was taken up indichloromethane (15 mL). At which point, pyridine (32 mL, 0.39 mmol) wasadded. The solution was stirred for 5 minutes, at which time, propionylanhydride (51 mL, 0.40 mmol) was added. The reaction was allowed to stirat room temperature for 72 h. An additional propionyl anhydride (0.1 eq)was added and the reaction monitored until complete disappearance ofstarting material was observed (MS, reaction may take up to 1 week). Thereaction was washed successively with a saturated aqueous solution ofNH₄Cl (3×) and H₂O (3×). The organic phase was dried over anhydrousNa₂SO₄ and evaporated in vacuo. Co-evaporation with toluene is necessaryto remove residual pyridine from the system. This was followed byre-dissolving the residue in DCM and solvent evaporation twice toproduce E-5 as a white foam (210 mg, 64%).

Method 3: A-1 (300 mg, 0.40 mmol), was dissolved in DCM (10 mL); to thissolution was added TEA (279 μL, 5 eq) and propionylchloride (175 μL, 5eq) subsequently and the mixture was stirred overnight at roomtemperature. Additional TEA (112 μL, 2 eq) and propionyl chloride (70μL, 2 eq) were added and again mixture was stirred overnight at roomtemperature. Once more additional TEA (112 μL, 2 eq) and propionylchloride (70 μL, 2 eq) were added and stirring at room temperature wascontinued overnight. TEA (344 μL, 9 eq) and propionyl chloride (314 μL,9 eq) were added and the mixture was stirred at room temperature for 5days. The reaction mixture was washed with aqueous Na₂CO₃-solution (3×,10%) and water (3×), dried, concentrated to dryness, and dried at theoil pump. ESI-MS (positive) showed tri- and tetra-propionylation.

Method 4

Propionic acid (4 eq) was taken up in 5 mL dichloromethane (DCM).Compound A-16 (0.5 mmol) and 4-dimethylaminopyridine (DMAP) (4.4 eq)were added and the resulting solution was cooled in an ice bath forapproximately 10 minutes. At this point, dicyclohexylcarbodiimide (DCC)(4.4 eq) was added slowly. The reaction was stirred continually at thistemperature for 5 minutes and then progressively warmed to roomtemperature where it was stirred overnight. Dicyclohexylurea (DCC) thatwas formed during the reaction is filtered off and discarded. Thefiltrate was collected and then washed with a saturated solution ofsodium hydrogencarbonate (3×), water (1×) and dried over anhydrousNa₂SO₄. The solvent was evaporated in vacuo. This was followed byre-dissolving the residue in a small volume of methanol. The solutionwas transported dropwise into ice-cold water (2× volume of methanol) andstored in the freezer overnight. The precipitated product was filteredoff and dried under high-vacuum to produce

TABLE 4 Propionylation Examples Reaction MS Compound SynthesisSubstituent Condition Degree of m/z Entry Method ALC equivalent (i.e.Workup) Substitution Yield ([M + H]⁺) E-5 1 A-1 1.1 as described 1 64%850.3 above  E-18 See A-1 N/A 1 80% 805.5 example 21  E-419 4  A-16 4 150% 791.3  E-420 2 (based on 847.1  E-421 3 tri-ester 903.0  E-52 3 A-118 3 Not 917.9  E-51 4 determined 973.8

Example 6

Synthesis of E-8

Erythromycin oxime was nitrated as described in the general procedure,followed by subsequent acetylation of crude nitration product withacetic anhydride in DCM with pyridine as catalyst. Column chromatography(acetone-cyclohexane 1:3) furnished the product E-8 as white amorphousfoam (39%, two steps).

Example 7

Synthesis of E-10

A solution of E-9 (1 mmol) in MeOH (20 ml) was vigorously stirred at 50°C. until TLC (acetone-cyclohexane 1:3) indicated complete deacetylationof the starting materials. Volatile components were removed in vacuo,the product E-10 was obtained as white amorphous foam (81%).

Example 8

Synthesis of E-11

Azithromycin was nitrated as described above. After desiredmono-nitration MeOH was added to the reaction mixture and stirring wascontinued for one further hour. Thus in situ generated methyl nitrateacted as methylating agent transferring one methyl group to11-O-position of the macrolide at ambient temperature. Standard aqueousworkup with subsequent purification by column chromatography(acetone-cyclohexane 1:3) delivered methylated macrolide nitrate E-11 aswhite amorphous solid (63%, two steps).

Example 9

Synthesis of E-16

Azithromycin (20.0 g; 26.7 mmol) was dissolved in 120 ml of MeOH. NaHCO₃(6.0 g; 71.5 mmol) was added, followed by a solution of K₂CO₃ (12.0 g;in water (80 ml; cooled down to RT), and finally iodine (6.3 g; 24.8mmol). The mixture was stirred vigorously at ambient temperature untilthe dark color had disappeared. A second batch of iodine (6.3 g; 24.8mmol) and K₂CO₃ carbonate (4.2 g; 30 mmol) were added. The procedure[addition of iodine 6.3 g and K₂CO₃ (4.2 g; 30 mmol)] was repeated untilMS showed (almost) full conversion. Sodium bisulfite was added to removeexcess oxidants, and all volatiles were evaporated. The solid residuewas finely ground and extensively extracted via Soxhlet extraction withacetonitrile. The extract was concentrated to ca. 75 ml and leftstanding at ambient temperature at least for 1 day and subsequently foranother day in the fridge. All solids were collected and recrystallizedfrom MeOH spiked with ca. 1 to 2 ml of water. Crystallization proceededfor about 3 days in an open vessel to yield 10 g (51%) of3′-N-demethyl-azithromycin (E-16). A second crop can be obtained fromthe mother liquors.

Example 10

Synthesis of E-20

A solution of E-16 (5 mmol) in DMSO (20 ml) was treated with decylbromide (6 mmol) at ambient temperature. Stirring continued for 12 huntil TLC (acetone-cyclohexane 1:3, 1% Et₃N) indicated consumption ofstarting materials. The system was diluted with EtOAc. (50 ml) andextracted with water (3× 30 ml). The organic phase was dried over sodiumsulfate. Evaporation of the solvent and drying at vacuum yielded E-20 aswhite amorphous foam (61%).

Example 11

Synthesis of E-25

A solution of azithromycin (13 mmol) and pyridine (13 mmol) in DCM (80ml) was cooled to 0° C. in an ice bath. A solution of acetic anhydride(14 mmol) in DCM (20 ml) was slowly added to the system. Afterwards thereaction mixture was allowed to warm up to ambient temperature andstirring was continued until TLC (acetone-cyclohexane 1:3, 1% Et₃N)indicated complete consumption of the starting materials. The system wasextracted with an aqueous solution of NH₄Cl (2× 50 ml) and water (3× 50ml). After drying over Na₂SO₄ all volatile components were evaporated invacuo, traces of pyridine were removed by coevaporation with toluene(2×). The product E-25 was a white amorphous solid (67%).

Example 12

Synthesis of E-22

A solution of E-25 (4 mmol) in dry THF (30 ml) was cooled to 0° C. in anice bath. A solution of propargyl bromide (4.4 mmol, 80% in toluene) wasadded slowly to the system. Afterwards the reaction mixture was allowedto warm up to ambient temperature and stirring was continued until TLC(acetone-cyclohexane 1:3, 1% Et₃N) indicated complete consumption of thestarting materials. The system was diluted with EtOAc (50 ml) andextracted with water (3× 50 ml). After drying over Na₂SO₄ volatilecomponents were evaporated in vacuo. Purification by columnchromatography (acetone-cyclohexane 1:3, 1% Et₃N) furnished E-22 aswhite amorphous powder (57%).

Example 13

Synthesis of E-12

A solution of E-22 (0.5 mmol), I-5 (0.5 mmol) and DIPEA (1 mmol) intoluene (5 ml) was treated with triethylphosphito copper(I) iodidecomplex (0.05 mmol) at ambient temperature. Stirring was continued untilTLC (ethyl acetate-cyclohexane 1:1) indicated complete consumption ofthe starting materials. The mixture was concentrated in vacuo andpurified by column chromatography (acetone-cyclohexane 1:1→acetone). Theproduct E-12 was obtained as colorless foam (32%).

Example 14

Synthesis of E-23

A solution of E-25 (5 mmol) and pyridine (5 mmol) in DOA (40 ml) wastreated with acetic anhydride (7 mmol) at ambient temperature. Stirringwas continued until TLC (acetone cyclohexane 1:3, 1% Et₃N) indicatedcomplete consumption of the starting materials. The system was extractedwith an aqueous solution of NH₄Cl (2× 20 ml) and water (3× 30 ml). Afterdrying over Na₂SO₄ all volatile components were evaporated in vacuo,traces of pyridine species were removed by co-evaporation with toluene(2×). The product E-23 was a white amorphous powder (54%).

Example 15

Synthesis of E-17

A solution of E-23 (2 mmol) in MeOH (30 ml) was vigorously stirred at50° C. until TLC (acetone-cyclohexane 1:3, 1% Et₃N) indicated completedeacetylation of the starting materials. Volatile components wereremoved in vacuo, the product E-17 was obtained as white amorphous foam(73%).

Alternative Synthesis: Compound E-2 (619 mg, 0.74 mmol) was charged in around bottom flask. To this was added a solution of acetic acid/methanol(2:1, 15 mL). To the stirred solution, was added Zn powder (368 mg, 5.63mmol, 7.6 eq). The resulting suspension was stirred at room temperatureand progressively monitoring the disappearance of the starting material(approx. 3 h). The suspension was filtered and the filtrate evaporatedin vacuo. The residue was taken up in dichloromethane (15 mL) producingsome white precipitate. The precipitate was filtered off. Thedichloromethane filtrate was washed with 10% aqueous Na₂CO₃ solution(2×), water (1×) and dried over anhydrous Na₂SO₄. The solvent wasevaporated in vacuo producing E-17 as a white solid (475 mg, 81% yield).

Example 16

Synthesis of E-26

A solution of azithromycin (13 mmol) and pyridine (13 mmol) in DCM (80ml) was cooled to 0° C. in an ice bath. A solution of benzoyl chloride(14 mmol) in DCM (20 ml) was slowly added to the system. Afterwards thereaction mixture was allowed to warm up to ambient temperature andstirring was continued until TLC (acetone-cyclohexane 1:3, 1% Et₃N)indicated complete consumption of the starting materials. The system wasextracted with an aqueous solution of NH₄Cl (2× 50 ml) and water (3× 50ml). After drying over Na₂SO₄ all volatile components were evaporated invacuo, traces of pyridine species were removed by co-evaporation withtoluene (2×). Column chromatography (acetone-cyclohexane 1:3, 1% Et₃N)yielded the product E-26 as a white amorphous foam (44%).

Example 17

Synthesis of E-13

A solution of E-5 (1 mmol) and propionic acid (1.2 mmol) in1,2-dichloroethane (3 ml) was treated portion wise with EDCI (3× 0.8mmol) and DMAP (1 mmol). The mixture was heated to 55° C. and stirredfor 6 days. After TLC (acetone-cyclohexane 1:3) indicated completeconsumption of the starting materials the mixture was diluted with EtOAc(30 ml) and extracted with water (3× 20 ml). After drying over Na₂SO₄the organic phase was evaporated and the remains were purified by columnchromatography (acetone-cyclohexane 1:3). The product E-13 was obtainedas colorless amorphous foam (42%).

Example 18

Synthesis of E-14

3′-N-desmethyl-azithromycin (I-1) (300 mg; 0.41 mmol) and1-bromo-3-nitrooxy-propane^([17] ()90 mg, 0.49 mmol) were dissolved indry DMSO (1.2 ml). The mixture was shaken at 23° C. for 3 hours.Afterwards additional 1-bromo-3-nitrooxy-propane (90 mg, 0.49 mmol) wasadded and shaking was continued for another hour. Once again,1-bromo-3-nitrooxy-propane (90 mg, 0.49 mmol) was added, the reactionmixture was shaken for additional 90 min., and then kept in the freezer(−16° C.) over night. The next morning additional1-bromo-3-nitrooxy-propane (90 mg, 0.49 mmol) was added and the mixturewas shaken for one hour. Water and DCM were added; after extraction, theorganic phase was dried (Na₂SO₄) and concentrated to dryness (withoutheating). The crude product was purified by column chromatography(eluent: CHCl₃:Isopropanol: NH₃ (7 M in MeOH 30:1:1).

Example 19

Synthesis of (I-5)

Azido-β-D-Glucopyranoside is synthesized from corresponding sugaracetate as is known to literature^([16]). After removal of anyprotective groups sugar azide is nitrated following above procedure. Dueto its high explosive risk the substance is always kept as DCM solutionand is stashed in the refrigerator.

Example 20

Synthesis of E-24

Compound E-1 (405 mg, 0.51 mmol) was taken up in dichloromethane (15mL). The resulting solution was cooled to 0° C. After 5 minutes, butyrylchloride (60 mL, 61.8 mg, 0.58 mmol, 1.1 eq.) was added. The reactionwas stirred for 10 min. at this temperature, at which point, thereaction was allowed to warm to room temperature, where it was stirreduntil reaction completion (2 h, or upon continuous monitoring). Thereaction was washed with a 10% Na₂CO₃ aq. solution (3×) followed by H₂O(3×), dried over anhydrous Na₂SO₄ and evaporated in vacuo to give E-24as a white foam (332 mg, 75% yield).

Example 21

Synthesis of E-18

Compound E-5 (CSY 1076) (126 mg, 0.15 mmol) was charged in a roundbottom flask. To this was added a solution of acetic acid/methanol (2:1,12 mL). To the stirred solution, was added Zn powder (74 mg, 1.12 mmol,7.6 eq). The resulting suspension was stiffed at room temperature andprogressively monitoring the disappearance of the starting material(approx. 3 h). The suspension was filtered and the filtrate evaporatedin vacuo. The residue was taken up in dichloromethane (10 mL) producingsome white precipitate. The precipitate was filtered off. Thedichloromethane filtrate was washed with 10% aqueous Na₂CO₃ solution(2×), water (1×) and dried over anhydrous Na₂SO₄. The solvent wasevaporated in vacuo producing E-18 as a white solid (96 mg, 80% yield).

The reduction conditions for removal of the NO₂ group was applied to thesyntheses of E-19 from E-24 providing the desired product in 78% yield.

Example 22

Synthesis of E-19

Compound E-24 CSY 4636 (100 mg, 0.12 mmol) was charged in a round bottomflask. To this was added a solution of acetic acid/methanol (2:1, 12mL). To the stirred solution, was added Zn powder (88 mg, 1.35 mmol,11.6 eq). The resulting suspension was stirred at room temperature andprogressively monitoring the disappearance of the starting material(approx. 3 h). The suspension was filtered and the filtrate evaporatedin vacuo. The residue was taken up in dichloromethane (15 mL) producingsome white precipitate. The precipitate was filtered off. Thedichloromethane filtrate was washed with 10% aqueous Na₂CO₃ solution(2×), water (1×) and dried over anhydrous Na₂SO₄. The solvent wasevaporated in vacuo producing E-19 as transparent gel (74 mg, 78%yield).

Example 23

Synthesis of E-81

250 mg of 2′-(2-Mercaptoethoxy)carbonyl-3-decladinosylazithromycin and250 mg of ammonium polysulfide are mixed with 10 ml of degassed andargonized glacial acetic acid and stirred with exclusion of oxygen for12 h. All volatiles are removed in vacuo, and the residue is extractedwith oxygen free saturated aqueous sodium hydrogen carbonate solution 3times. The residue is washed with water (oxygen free), dried in vacuoand used as such.

Example 24

Synthesis of E-82

250 mg of 2′-(2-Mercaptoethoxy)carbonyl-3-decladinosylazithromycin aredissolved in a mixture of 5 ml of tert. butanol and 5 ml ofdichloromethane. 250 μl of tert. butylnitrite are added, and the mixtureis stirred for 48 h with exclusion of light. All volatiles are removedi.v. keeping the temperature below 20° C. and light excluded. The redresidue is used as such

Example 25

Synthesis of E-74

380 mg of Azithromycin are dissolved with 20 ml of DMF and cooled in anice bath. 41 ml of pyridine and then 85 mg (1.1 eq.) of rac-2-acetoxypropionyl chloride, dissolved with 1 ml of dichloromethane, are addedand the mixture is allowed to warm up to room temperature with stirring.When mass spectrometry indicates consumption of the macrolide, themixture is diluted with 50 ml of ethyl acetate, extracted twice withwater, 3 times with saturated aqueous sodium hydrogen carbonatesolution, once again with water and brine, each, and dried over sodiumsulfate. After evaporation and vacuum drying, the diastereomeric mixture(1:1) of target compound E-74 remains as a slightly yellowish foam.

Yield: 385 mg

MS: m/z=863.5 ([M+H]⁻)

The same procedure can be applied in preparing E-77, E-83, E-84, E-85and E-86.

TABLE 5 Typical Products from the Acylation Procedures Entry Acylatingagent Product structure Yield [%] ([M + H]⁺ E-83 O-Phenylchlorothionoformate

66 885.7 E-85 2- Bromoethylchloroformate

82 899.6 E-84 Hexanoylchloride

93 847.6 E-77 Lipoyl chloride

72 936.8 E-86 Allylchloroformate

82 833.5 E-540 O-Phenyl chlorothionoformate

23 884 E-26 Benzoyl

44 853

Example 26

General Procedure: Synthesis of n-Butyryl-Propanolol, Compound E-133

Propranolol HCl (200 mg, 0.68 mmol) was taken up in dichloromethane (4mL). To this was added dropwise butyryl chloride (69 μL, 0.71 mmol) andthe reaction was stirred at room temperature for 1 hour. To the reactionwas added triethylamine (194 μL, 1.4 mmol). After 30 min, additionalbutyryl chloride (30 μL, 0.3 mmol) was added. Reaction was monitored bythe disappearance of starting material. The reaction was stopped after20 min by the addition of 10 mL 10% aqueous Na₂CO₃ solution. The twophases were stirred for 5 min separated. The organic layer was washedsuccessively with 10% aqueous Na₂CO₃ (1×), H₂O (1×) and saturated aq.NaCl (1×), dried with Na2SO4 and evaporated in vacuo to get an oil film.5 mL HCl in Et₂O (2M) and 1 mL MeOH was added and stirred for 5 min,then evaporated in vacuo and dried with the vacuum pump under nitrogenfor 1 hour to get a brown oil (yield 91%).

Example 27

General Procedure: Synthesis of n-Butyryl-Hydroxychloroquine, CompoundE-89

Hydroxychloroquine sulfate (1099 mg, 2.53 mmol) was charged into a roundbottom flask. H₂O (10 mL) and dichloromethane (10 mL) were added.Pyridine (412 μL, 5.1 mmol) was added and the reaction stirredvigorously for 5 min. Butyric anhydride (420 μL, 2.65 mmol) was addedand the reaction stirred at room temperature for 3 h. The phases wereseparated and the dichloromethane layer was washed successively with asaturated aqueous NH₄Cl solution (2×15 mL), H₂O (2×10 mL), dried overNa₂SO₄ and evaporated in vacuo. Co-evaporation with toluene is necessaryto remove residual pyridine from the system. This was followed byre-dissolving the residue in DCM and solvent evaporation twice toproduce a yellow oil (93 mg, 9% yield).

This procedure can be applied for the synthesis of the followingcompounds E-87 to E-88 E-90 to E-97.

Example 28

Typical methylation reaction of Hydroxychloroquine or Propranolol wasachieved using Eschweiler-Clarke-methylation reaction. Acylationreactions were carried out using typical procedures described above.

Example 29

Compounds, prepared by reacting a carrier molecule with acylatingagents.

These carrier molecules are prepared by reacting symmetric orunsymmetric di- or poly-epoxides with secondary amines, thus containingthe common structural element of 2 or more alcohols, vicinallyneighbored by a tertiary amine.

Alternatively, carrier molecules can be prepared by reacting epoxideswith diethanolamine. The reaction products are containing 2-hydroxytertiary amines.

TABLE 6 Examples for di- or polyepoxides No. of epoxide Entry NameCAS-No. functions A 1,3-Butadiene diepoxide 1464-53-5 2 B 1,4-Butandiolediglycidylether 2425-79-8 2 C N,N-Diglycidylaniline 2095-06-9 2 DResorcinol diglycidylether 101-90-6 2 E Ethylen glycol diglycidylether2224-15-9 2 F 1,7-octadienediepoxide 2426-07-5 2 G Diglycidyl ether2238-07-5 2 H 1,2,4,5,9,10-Triepoxydecane 52338-90-6 3 1N,N-Diglycidyl-4-glycidyloxyaniline 5026-74-4 3 J Poly(ethylene glycol)diglycidyl ether 72207-80-8 2 (average M_(n) 500) K Glyceroldiglycidylether 72207-80-8 2 L 4,4′-Methylenebis(N,N- 28768-32-3 4diglycidylaniline) M Bis[4-(glycidyloxy)phenyl]methane 2095-03-6 2

TABLE 7 Examples for secondary amines Entry Name CAS-No. 1 Dimethylamine2 Morpholine 3 4-Methylpiperazine 4 Piperidine 51,2,3,4-Tetrahydroisoquinoline 91-21-4 6 Diethylamine 7 Dioctylamine 8Diethanolamine 9 Sarcosine methyl ester hydrochloride 13515-93-0 10(R)-pyrrolidine-2-carboxylic acid methyl ester (Prolin methylester) 11Ethyl 1,4-diazepan-1-ylacetate dihydrochloride

TABLE 8 Structural examples for carrier molecules Combination ofpoly-epoxide and amine Structural Example H1

G5

C8

A-7-C

B2

The 2-aminoalcohols of these carriers can be esterified to short chaincarboxylic acids or nitric acid. One molecule can contain esters ofdifferent of these acids. Examples for short chain carboxylic acids are:

Acetic acid, propionic acid, butyric acid, isobutyric acid, valericacid, 2-methylbutyric acid, 3-methylbutryric acid, lactic acid, pyruvicacid, 3-phenylpropionic acid, succinic acid, maleic acid, fumaric acid,malic acid, lactic acid butyrate (lactic acid butanioate), 2-acetoxypropionic acid, mandelic acid, benzoic acid.

Structural Examples of such Esters are:

Alternatively, these carrier molecules are prepared by reactingsymmetric or unsymmetric di- or polyamines with epoxides, thuscontaining the common structural element of 2 or more alcohols,vicinally neighboured by a tertiary amine.

TABLE 9 Examples for di- or polyamines No. of reactive Entry Name CAS.No. amines N Spermine 71-44-3 4 O Spermidine 124-20-9 3 P Piperazine110-85-0 2 Q 1-(2-Aminoethyl)piperazine 140-31-8 2 R L-Lysine 56-87-1 2S Homopiperazine 505-66-8 2 T 1,3-Diamino-2-propanol 616-29-5 2 U1,3,5-Triamino-1,3,5-trideoxy-cis-inositol 6988-69-8 3 trihydrochlorideV 1,2,3,4-Tetrahydroquinoxaline 3476-89-9 2 W Tetraethylenepentamine112-57-2 5

TABLE 10 Examples for epoxides Entry Name CAS No. 12 Propylene oxide75-56-9 13 Cyclohexen oxide 286-20-4 14 Ethyl 2,3-epoxypropionate4660-80-4 15 Styrene oxide 96-09-3 16 Glycidol 556-52-5

TABLE 11 Structural examples for carrier molecules Combination ofpoly-epoxide and amine Structural Example Q12

Q13

R16

S14

T15

The 2-aminoalcohols of these carriers can be esterified to short chaincarboxylic acids or nitric acid. One molecule can contain esters ofdifferent of these acids. Examples for short chain carboxylic acids are:

Acetic acid, propionic acid, butyric acid, isobutyric acid, valericacid, 2-methylbutyric acid, 3-methylbutryric acid, lactic acid, pyruvicacid, 3-phenylpropionic acid, succinic acid, maleic acid, fumaric acid,malic acid, lactic acid butyrate (lactic acid butanoate), 2-acetoxypropionic acid, mandelic acid, benzoic acid.

Structural Examples of such Esters are:

Example 30

Synthesis of Rac. 2′-Deoxy-2′-S-Thioacetyl ppropranolol^([18])

Diethyl azodicarboxylate (DEAD, 10 mmol) is added to a stirred (magneticstirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (25mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol)and thioacetic acid (10 mmol) both dissolved in THF (10 mL) are addeddropwise and stirring is continued for 1 h at 0° C. and further 2 h atambient temperature. Any precipitates are filtered off, the remainingsolution is concentrated in vacuo and desired product is isolated bycolumn chromatography (silica gel, cyclohexane-ethyl acetate).

Synthesis or Rac. 2′-Deoxy-2′-S-Thio Propranolol Sodium Salt

rac. 2′-Deoxy-2′-S-thioacetyl propranolol (10 mmol) is dissolved inmethanol (20 mL) while stirring (magnetic stirrer, 300 rpm) and sodiummethoxide (10 mmol) is added at 0° C. The system is allowed to warm upto ambient temperature and treatment is continued until TLC(cyclohexane-ethyl acetate) indicates full conversion of startingmaterials. Afterwards the reaction mixture is rinsed into ice colddiethyl ether (100 mL). Any precipitates are filtered off and are driedin vacuo to yield a white to slightly yellow product.

Example 31

Synthesis of Rac. 2-O-Nitrolactic acid

Lactic acid (10 mmol) is suspended in acetonitrile (20 mL) in a 3-neckedround bottom flask and is cooled to 0° C. in an ice bath while stirring(300 rpm). Diphosgene is added (5 mmol) followed by careful dropwiseaddition of silver nitrate solution (20 mmol, dissolved inacetonitrile). The mixture is stirred for 30 min at 0° C., subsequentlyis allowed to warm up to ambient temperature and stirring is continuedfor further 30 min. Afterwards any precipitates are filtered off and themixture is carefully concentrated in vacuo, As crude products are likelyto be explosive compounds the system was not fully dried but taken up inTHF (10 mL) to be immediately used in the following step.

Synthesis of Rac. 2′-O-(2-O-Nitrolactyl) Propranolol—E-42

Diethyl azodicarboxylat (DEAD, 10 mmol) is added to a stirred (magneticstirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (25mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol)and rac. 2-O-nitrolactic acid (10 mmol) both dissolved in THF (10 mL)are added dropwise and stirring is continued for 1 h at 0° C. andfurther 2 hours at ambient temperature. Any precipitates are filteredoff, the remaining solution is concentrated in vacuo and desired productis isolated by column chromatography (silica gel, cyclohexane-ethylacetate).

Example 32

Rac. Bis-(2′-Deoxy-2′-S-S-Disulfido Propranolol) (Putative)^([19])

A round bottom flask is charged with ethyl acetate (10 mL), graphite (3g), iodine (0.5 mmol) and cerium(III) chloride heptahydrate (1 mmol).The mixture is stirred for 10 min at ambient temperature, followed byaddition of rac. 2′-deoxy-2′-S-thio propranolol sodium salt (10 mmol).Treatment is continued until TLC (cyclohexane-ethyl acetate) indicatesfull conversion of starting materials. After completion the system isfurther diluted with additional ethyl acetate (250 mL) and is washedwith a saturated solution of aqueous sodium thiosulfate, water, and isdried over sodium sulfate. Upon filtration any volatiles are removed invacuo and the residue is subject to column chromatography (silica gel,cyclohexane-ethyl acetate).

Mixed disulfides may also be accessible in this way but require slightalterations.

Example 33

Typical Example or Synthesizing ALC Cores A-8 and A-9

Synthesis of 2-(4-Pyridyl)-3-Amino-4-(3-Methoxyphenyl)Carbonyl Pyrazole

10.96 g of 4-(N-phenyl)amino-3-(3-methoxyphenyl)carbonylacrylonitrileand 6.27 g of 4-pyridylhydrazine hydrochloride are combined with 6.2 mlof triethylamine in 140 ml of ethanol, flushed with argon and heated toreflux for 5 h. The mixture is concentrated to 50 ml and diluted with200 ml of cyclohexane. The precipitate is filtered off and washed withdiethyl ether, until no further colour is extracted any more. Theremaining solid is dissolved with a mixture of dichloromethane and water(300 ml each). The organic phase is dried with brine and sodium sulfateand concentrated to dryness.

Yield: 7.07 g (61%); MS: m/z=295 ([M+H]+)

Synthesis of 2-(4-Pyridyl)-3-Amino-4-(3-Hydroxyphenyl)Carbonyl Pyrazole

2.6 g of 2-(4-Pyridyl)-3-amino-4-(3-methoxyphenyl)carbonylpyrazole aresuspended in 10 ml of a solution of 33% hydrobromic acid in acetic acid.The mixture is heated to 70° C. for 16 h. After cooling, the reactionmix is poured into 150 ml of water. The precipitate is filtered off,washed with saturated aqueous sodium hydrogen carbonate solution (twice)and water, and dissolved in 10 ml of a solution of ammonia in methanol(7M). After 30 min, all volatiles are removed by evaporation, and theresidue is dissolved in 50 ml of boiling methanol. The product isprecipitated by pouring into 250 ml of water. Filtration and dryingyield 2.35 g of an off white powder.

Synthesis of2-(4-Pyridyl)-3-Amino-4-(3-[2,3-Dihydroxypropyloxy]Phenyl)-CarbonylPyrazole

2.09 g of 2-(4-Pyridyl)-3-amino-4-(3-hydroxyphenyl)carbonyl pyrazole aredissolved with 25 ml of dimethylformamide. 3.5 g of potassium carbonateand 580 mg of glycidol are added. The mixture is kept stirring at 60° C.for 18 h. The reaction mixture is partitioned between water and ethylacetate. The organic phase is washed with water, dried with brine andsodium sulfate, and concentrated i.v. The residue is subjected topreparative HPLC to yield 1.4 g of the product.

Synthesis of2-(4-Pyridyl)-3-Amino-4-(3-[2,3-Di{Butyroyloxy}Propyloxy]Phenyl)CarbonylPyrazole E-199

500 mg of2-(4-pyridyl)-3-amino-4-(3-[2,3-dihydroxypropyloxy]phenyl)carbonylpyrazoleare dissolved with 5 ml of pyridine. 500 μl of butyric acid anhydrideare added, and the mixture is stirred at 50° C. over night. 1 ml ofmethanol is added, and the mixture is stirred for further 30 min. Thereaction is allowed to reach room temperature and partitioned betweenwater and ethyl acetate. The organic phase is extracted with water 5times, then with brine and dried over sodium sulfate. After evaporationof all volatiles, the product is purified by chromatography over silicagel.

Yield: 290 mg

The same conditions can be applied to synthesis of2-(4-fluorophenyl)-3-amino-4-(3-[2,3-di{butyroyloxy}propyloxy]phenyl)carbonylpyrazoleE-210

Example 34

Synthesis of Chenodeoxycholic Acid Azithromycin-2′-Ester

1 g of Chenodeoxy cholic acid is dissolved with 50 ml of drydichloromethane and cooled in an ice bath. 500 mg of carbonyldiimidazole are added, and the mixture is stirred for 2 h, whilereaching room temperature. 2 g of Azithromycin are added, and themixture is stirred for 72 h. The mixture is extracted with water (3×)and then with 5% citric acid. The citric acid phase is washed withdichloromethane (2×). It is then vigorously stirred with ethyl acetate,while portions of sodium hydrogencarbonate are added, so that gasevolution is under control. When no gas is developed any more, theorganic phase is isolated, washed with brine and dried over sodiumsulfate. Concentration and chromatography with a gradient starting at10% of acetone in cyclohexane (always containing 0.2% of triethylamine)yields 350 mg of the desired product.

Example 35

Synthesis of2′-(Succinyl-1-Hydroxymethylferrocene)-11-Nitro-Azithromycin

Compound E-10 (200 mg, 0.25 mmol) was dissolved in dry dichloromethane(5 mL). To this was added subsequently, 4-dimethylaminopyridine (4-DMAP,3 mg, 0.25 mmol. 0.1 eq.) and succinic anhydride (28 mg, 0.28 mmol). Thereaction was stirred overnight at room temperature. The solvent wasremoved in vacuo and the resulting white amorphous foam was useddirectly for the next step.

Fresh dry dichloromethane (5 mL) was added to the resulting foam,followed by 1-Hydroxy-methylferrocene (60 mg, 0.28 mmol, 1.1 eq.). Thereaction was cooled to 0° C. and to this was added EDCI (96 mg, 0.5mmol, 2 eq.). The reaction was allowed to progressively warm to roomtemperature where it was stirred overnight. Additional dichloromethane(20 mL) was added and washed several times with saturated aqueousammonium chloride, brine (2×), dried under anhydrous Na₂SO₄ and thesolvent removed in vacuo. The resulting crude product was purified bychromatography with a gradient starting at 10% of acetone in cyclohexane(0.2% Et₃N) yields 150 mg of the desired product (53%).

Similarly, the following compound may be obtained using the procedureabove starting from compound E-19

Example 36

Activity of substances in the inhibition of growth of bacteria. Bacteriaincluding the species Escherichia coli, Bacillus pumilus, Salmonellasp., Micrococcus luteus and Staphylococcus carnosus are cultured inappropriate media (Luria broth for all except S. canosus). Overnightcultures are mixed with fresh medium to reach an optical density at 600nM of ca. 0.1 AU. These cultures are mixed with solutions of substancesto be tested at concentrations ranging from 100 μM to 0.05 μM in amicrotitre plate. The growth of the culture is monitored by measuringthe optical density at various times after the addition of theinhibitor. Reduction in the rate of increase in optical densitycorresponds to an inhibition of bacterial growth. In the followingtables, the activity of various of the test substances may be observedby reductions in optical density relative to untreated control cultures.The data are summarized in Table 3 and Table 4.

TABLE 12 Inhibition of growth of Staphylococcus carnosus by compoundsafter 9-20 h: Conc. Absorbance of culture medium at 600 nm (μM): 100 5025 13 6 3 2 0.8 E-2 0.395 0.425 0.456 0.542 0.626 0815 0.766 0.760 E-50.302 0.347 0.403 0.452 0.525 0.805 0.726 0.819  E-13 0.421 0.437 0.2820.488 0.530 0.668 0.755 0.765 E-1 0.157 0.096 0.078 0.068 0.625 0.8640.856 0.902  E-12 0.494 0.523 0.574 0.548 0.591 0.577 0.688 0.783 E-90.545 0.522 0.426 0.677 0.752 0.830 0.737 0.765  E-10 0.576 0.433 0.5950.702 0.699 0.768 0.826 0.862  E-11 0.641 0.574 0.819 0.822 0.887 0.8900.918 0.941 No 0.887 compound

TABLE 13 Inhibition of growth of Salmonella typhimurium by compoundsafter 9-20 h: Conc. Absorbance of culture medium at 600 nm (μM): 100 5025 13 6 3 2 0.8 E-2 0.251 0.168 0.314 0.621 0.382 0.410 0.441 0.452 E-50.390 0.395 0.396 0.437 0.478 0.511 0.568 0.574  E-13 0.398 0.407 0.4270.459 0.505 0.543 0.605 0.634 E-1 0.557 0.484 0.736 0.722 0.741 0.7110.761 0.722  E-12 0.332 0.270 0.326 0.343 0.400 0.354 0.457 0.455 E-90.221 0.319 0.395 0.382 0.348 0.327 0.272 0.300  E-10 0.280 0.315 0.3900.399 0.382 0.402 0.361 0.334  E-11 0.562 0.650 0.697 0.633 0.627 0.6230.587 0.555 No 0.943 compound

Example 37

Substances may act directly on bacteria, or they may act to promote thekilling of the bacteria by phagocytes. To measure this effect, culturesmurine macrophages are incubated with a test bacteria and the number ofbacteria surviving are counted in terms of the viable colony formingunits (CFU). The method for determining the rate of phagocytosis is asfollows:

Intracellular killing of S. Typhimurium by mouse macrophage cellline J774 A.1

seed a monolayer of cells in 200 μl Media into the wells of a 96 wellplates

incubate O/N 37° C.

remove medium and add fresh medium

add Bacteria (Salmonella typhimurium) e.g. 5 μl of 1:100 diluted O/Nculture (MOI=10) (=108 cfu/ml)

centrifuge 10 min 800 g (˜2000 rpm)

incubate 20-30 minutes at 37° C. (phagocytosis)

remove media

wash 1-2× with PBS

add medium with 100 μg/ml Gentamicin, stock: 10 mg/ml (=1:100)

incubate 45′ at 37° C.

wash 2× with PBS

add fresh medium (200 μl/well)

add compounds to test

incubate 2-3 hours at 37° C.

remove medium

lyse cells with water: add 200 μl H2O incubate 10′, push a few timesthrough 27 gauge needle using a 1 ml syringe

plate 100 μlf 1:10 dilution onto LB-agar plates (=1:100 dil)

Monolayer of J 774 A.1 in 96 well plate=˜1-5×104 cells

Overnight culture of Salmonella thyph.=˜1×1010 cfu/ml

Overnight culture of Staph. carnosus=˜5×109 cfu/ml MOI=50 (=5 μl of 1:10dil. O/N culture)

MOI=Multiplicity of Infection

Medium: DMEM/RPMI 7.5 FCS

Example 38

The potential efficacy of a Compound for Inflammatory bowel disease maybe modeled as follows. C57 BLK6 or BALBc mice are provided with drinkingwater containing 2.5% or 2.8% dextran sulfate. Animals are weighed andobserved for signs of intestinal disturbance daily. Signs includediarrhea or occult blood. Compound is formulated by mixing with asolution of 0.1 up to 1% citric acid depending on concentration.Compound is provided by oral gavage daily. Example data for the efficacyof compounds cited here is provided in FIG. 3, 7, 8 or 10-19 .

Example 39

The potential efficacy of a Compound for rheumatoid arthritis may bemodeled as follows. DBA1 mice are induced by a subcutaneous injection ofbovine collagen in 0.05M acetic acid, emulsified in Freund's adjuvant.21 days later, a second injection of this material is made withoutinclusion of mycobacterial material in the adjuvant. Animals are weighedand observed for signs of inflammation daily. Signs include weight loss,swelling of paws, redness and reduced mobility. Compound is formulatedby mixing with a solution of 1% citric acid. Compound is provided byoral gavage daily. Data for the efficacy of compounds cited here isprovided in FIG. 2 .

Example 40

The potential efficacy of a Compound in modulating immune reactions maybe determined as follows. Swiss or C57 Blk6 mice are induced to producecytokines by a subcutaneous injection of lipopolysaccharide, Typically,compound is provided at time 0. Compound is formulated by mixing with asolution of 1% citric acid for oral treatment or, dissolved in PEG 300and diluted in water for intra-peritoneal treatment. Compound isprovided by oral gavage. 30 minutes after providing compound, animalsare treated with an intra-peritoneal injection of a solution oflipopolysaccharide in the concentration range that will provide 0.01mg/kg lipopolysaccharide. Data for the efficacy of compounds cited hereis provided in FIG. 1 .

Example 41

The potential efficacy of a Compound in treating a malignant disease maybe determined as follows. Tumours are known to be deficient in nitricoxide and this is considered to be a cause of local tolerance. Providinga nitric oxide donor that is accumulated in macrophages in the tumourenvironment provides a means to artificially modify the local NO status.C57 Blk6 mice are injected subcutaneously with an murine ovarian cancercell line expressing ovalbumin. Mice bearing tumours are selected after14 days. Typically, compound is provided at this time. Compound isformulated by mixing with a solution of 1% citric acid for oraltreatment. Compound is provided by oral gavage. Animals are monitoreddaily for tumour size, body weight and activity score. The activity ofthe compound may be determined in combination with other therapiesincluding anti-bodies or vaccines based on a tumour antigen. In thiscase ovalbumin, can serve as a model antigen.

Example 42

Synthesis of 2′-O-(2-Ferrocenyl) Acetyl-11-O-Nitro-Azithromycin

11-O-Nitro-azithromycin (0.25 mmol) was dissolved in dry dichloromethane(5 mL). To this was added EDCI (2 eq., 0.5 mmol) and 2-ferrocenyl aceticacid (1.1 eq., 0.28 mmol). The reaction was stirred overnight at roomtemperature. The solvent was removed in vacuo and the resulting whiteamorphous foam. The resulting crude product was purified by columnchromatography with a gradient starting at 10% of acetone in cyclohexane(0.2% Et₃N).

Similarly, the following compound may be obtained using the procedureabove starting from 2′-O-Nitro-azithromycin:

2′-O-Nitro-11-O-(2-Ferrocenyl) Acetyl-Azithromycin

Example 43

Rac. 2′-O-Propionyl Propranolol

Diethyl azodicarboxylat (DEAD, 10 mmol) is added to a stirred (magneticstirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF(2.5 mL) at 0° C. and treatment is continued for 30 min. Propranolol (5mmol) and propionic acid (10 mmol) both dissolved in THF (10 mL) areadded dropwise and stirring is continued for 1 h at 0° C. and further 2hours at ambient temperature. Any precipitates are filtered off, theremaining solution is concentrated in vacuo and desired product isisolated by column chromatography (silica gel, cyclohexane-ethylacetate).

Example 44

Rac. 2′-O-Acetoxypropionyl Propranolol

Diethyl azodicarboxylate (DEAD, 10 mmol) is added to a stirred (magneticstirrer, 300 rpm) solution of triphenylphosphin (10 mmol) in dry THF (25mL) at 0° C. and treatment is continued for 30 min. Propranolol (5 mmol)and 2-acetoxypropionic acid (10 mmol) both dissolved in THF (10 mL) areadded dropwise and stirring is continued for 1 h at 0° C. and further 2hours at ambient temperature. Any precipitates are filtered off, theremaining solution is concentrated in vacuo and desired product isisolated by column chromatography (silica gel, cyclohexane-ethylacetate).

Example 45

Synthesis of Azithromycin 11,2′-Lipoate

380 mg of azithromycin-2′-lipoate (E-77) are dissolved in 25 ml ofdichloromethane and cooled in an ice bath. 125 mg of lipoyl are adde,then 50 μl of pyridine. The mixture is allowed to reach room temperatureand stirred for 16 h. The reaction mixture is extracted with water 3times, then once with 5% aqueous citric acid. The citric acid phase isextracted with dichloromethane, then combined with ethyl acetate andcarefully made basic with sodium hydrogen carbonate and vigorousstirring. When gas evolution ceases, the organic phase is separated,washed with water and brine, and dried with sodium sulfate. Afterevaporation of all volatiles, the residue is chromatographed with agradient starting at cyclohexane-acetone 5-1, containing 0.5% oftriethylamine. Yield: 140 mg

Example 46

Synthesis of Azithromycin 11-Lipoate

350 mg of Azithromycin 11,2′-dilipoate are stirred with 5 ml methanol atroom temperature. When mass spectrometry indicates completion of thereaction (m/z=1125.5→937.5), the mixture is partitioned between waterand ethyl acetate. The organic phase is washed once with water, thenextracted with 5% aqueous citric acid. The citric acid phase isextracted with dichloromethane, then combined with ethyl acetate andcarefully made basic with sodium hydrogen carbonate and vigorousstirring. When gas evolution ceases, the organic phase is separated,washed with water and brine, and dried with sodium sulfate. Afterevaporation of all volatiles, the residue is chromatographed with agradient starting at cyclohexane-acetone 5-1, containing 0.5% oftriethylamine. Yield: 225 mg

Example 47

Formation of Acetic Esters of ALCs

Method 1: ALC (1.0 mmol) was taken up in 15 mL dichloromethane. Pyridine(1.2 eq.) was added and the resulting solution was cooled in an ice bathfor approximately 10 minutes. At this point, a solution of aceticanhydride (1.2 eq) was added dropwise. The reaction was stirredcontinually at this temperature and then progressively warmed to roomtemperature where it was stirred overnight. Reaction progress wasmonitored either by TLC and/or MS. The reaction was washed with asaturated solution of ammonium chloride (3×), water (3×) and dried overanhydrous Na₂SO₄. The solvent was evaporated in vacuo. Co-evaporationwith toluene is necessary to remove residual pyridine from the system.This was followed by re-dissolving the residue in DCM and solventevaporation twice to produce a white foam, which was dried underhigh-vacuum to produce acetylated product.

This acetylation conditions can be extended for other ALCs. In the casewhere the acetylation proceeds sluggish, alternative reaction conditionswere undertaken as described below:

Method 2. Compound A-12 (0.85 mmol) was taken up in DCM (10 mL). To thiswas added triethylamine (3.5 eq) and acetyl chloride (3.5 eq). Reactionwas monitored by TLC and MS until disappearance of starting ALC.Reaction was filtered. The filtrate was either evaporated in vacuo anddirectly purified by column chromatography or the filtrate was washedwith 10% aq. Na₂CO₃ solution, brine, dried over Na₂SO₄ and evaporated invacuo to get the crude product.

Method 3. Acetic acid (4 eq) was taken up in 5 mL dichloromethane (DCM).Compound A-16 (0.5 mmol) and 4-dimethylaminopyridine (DMAP) (4.4 eq)were added and the resulting solution was cooled in an ice bath forapproximately 10 minutes. At this point, dicyclohexylcarbodiimide (DCC)(4.4 eq) was added slowly. The reaction was stirred continually at thistemperature for 5 minutes and then progressively warmed to roomtemperature where it was stirred overnight. Dicyclohexylurea (DCU) thatwas formed during the reaction is filtered off and discarded. Thefiltrate was collected and then washed with a saturated solution ofsodium hydrogencarbonate (3×), water (1×) and dried over anhydrousNa₂SO₄. The solvent was evaporated in vacuo. This was followed byre-dissolving the residue in a small volume of methanol. The solutionwas transported dropwise into ice-cold water (2× volume of methanol) andstored in the freezer overnight. The precipitated product was filteredoff and dried under high-vacuum to produce a product.

TABLE 14 Acetylation Examples Reaction MS Compound Synthesis SubstituentCondition Degree of m/z Entry Method ALC equivalent (i.e. Workup)Substitution Yield ([M + H]⁺) E-2 1 A-1 1.2 as described above 1 76% 837E-4 1 A-1 1.5 as described above 1 41% E-8 1 A-1 1.2 39%  E-23 1 A-1 2.0as described above 2 54%  E-25 1 A-1 1.1 as described above 1 67%  E-4183 A-16 4 as described above 3 20% CHMA02063  E-228 1 A-10 4 as describedabove 2 56% 673  E-453 A-17 Overall 1 and 2 49% 819.7 A-17 12 equiv.(referring to 861.5 Ac₂O the di-ester)   E-266/ 2 A-12 3 as describedabove, 2 > 4 77% 675, 759  E-268 filtered through a silica gel plugusing CHCl₃:iPrOH:7M NH₃ in MeOH (30:1:1) as mobile phase E-23/E-50 A-12 and 3 16% 833, 875  E-564 1 A-2 (X = O) 1 89% 776  E-29 1 E-19 1.2direct chromatography 1× 85% 861 for purification

Example 48

Formation of Butyric and Isobutyric Esters of ALCs

Method 1: Compound A-1 was taken up in dichloromethane and stirred for10 min. At this point, a solution of carboxylic anhydride andtriethylamine in dichloromethane was added dropwise. The reaction wasstirred continually at room temperature. The reaction solution waswashed with 5% citric acid three times to extract the product. Acidicsolution was then washed with ethyl acetate (2×) and afterwardsneutralized with Na₂CO₃. Product was extracted with ethyl acetate (3×).The solution was washed with a saturated solution of sodium chloride(2×), water (2×) and dried over anhydrous Na₂SO₄. The to solvent wasevaporated in vacuo to produce a white foam containing product.

Method 2: Compound A-1 was taken up in dichloromethane and was cooled inan ice bath for approximately 10 minutes. At this point, a solution ofcarboxylic chloride in dichloromethane was added dropwise. The reactionwas stirred continually at this temperature for 15 min and thenprogressively warmed to room temperature where it was stirred for 2.5 h.

The reaction was washed with a 10% solution of Na₂CO₃ (3×), water (3×)and dried over anhydrous Na₂SO₄. The solvent was evaporated in vacuo.Co-evaporation with toluene is necessary three times. This was followedby re-dissolving the residue in dichloromethane to produce a white foam,which was dried under high-vacuum to produce product.

Method 3: Starting material was taken up in dichloromethane and stirredfor 10 min. At this point, a solution of carboxylic chloride andtriethylamine in dichloromethane was added dropwise. The reaction wasstirred continually at room temperature for two days. The reactionsolution was washed with 5% citric acid three times to extract theproduct. Acidic solution was then washed with ethyl acetate (2×) andafterwards neutralized with Na₂CO₃. Product was extracted with ethylacetate (3×). The solution was washed with a saturated solution ofsodium chloride (2×), water (2×) and dried over anhydrous Na₂SO₄. Thesolvent was evaporated in vacuo to produce a white foam containingproduct.

Method 4: Compound E-48 or E-39 was solved in methanol to hydrolyzebutyric esters. The reaction was stirred continually at room temperaturefor two days. The reaction solution was washed with ethyl acetate threetimes to extract the product. The ethyl acetate phase was washed with 5%citric acid (3×). Acidic solution was then washed with ethyl acetate(2×) and afterwards neutralized with Na₂CO₃. Product was extracted withethyl acetate (3×). The solution was washed with a saturated solution ofsodium chloride (2×), water (2×) and dried over anhydrous Na₂SO₄. Thesolvent was evaporated in vacuo to produce a white foam containingproduct.

Method 5: Carboxylic acid (180 mg, 2.04 mmol) was solved in 3 mLdichloromethane. Under stirring conditions 4-Dimethylaminopyridine (274mg, 2.24 mmol) and A-16 were added. The reaction solution was cooled to0° C. and N,N′-Dicyclohexylcarbodiimide (463 mg, 2.24 mmol) was added.The reaction was stirred continually at this temperature for 5 min andthen progressively warmed to room temperature where it was stirred for12 h. Precipitation was removed via filtration. The reaction was washedwith a saturated solution of NaHCO₃ (3×) and dried over anhydrousNa₂SO₄. The solvent was evaporated in vacuo. Product was solved inmethanol and water was added. Solution was cooled to −20° C., aprecipitation occurred which was extracted and dried in vacuo.

Workup 1: Column chromatography over silica gel was carried out toseparate different products. As eluent a mixture of chloroform,2-propanol and ammonia in methanol (60:1:1) was used. The solvent wasevaporated in vacuo.

Workup 2: Preparative chromatography over RP-C18-silica gel was carriedout to separate different products. As eluent a mixture of water (withtrifluoroacetic acid 0.05%) and methanol (with trifluoroacetic acid0.05%) was used. The solvent was evaporated in vacuo.

Workup 3: Column chromatography over silica gel was carried out toseparate different products. As eluent a mixture of cyclohexane, acetone(7:1) with 0.5% triethylamine was used. The solvent was evaporated invacuo.

TABLE 15 Butyrylation/Isobutyrylation Examples Compound SynthesisReaction Condition Degree of Entry Method ALC Substituent (i.e. Workup)Substitution Yield MS E-35 1 A-1 Butyric Workup 1 1x 76% 819 E-39 3 A-1Butyric Workup 2 2x 10% 889 E-48 3 A-1 Butyric Workup 2 3x 12% 959 E-473 A-1 Butyric Workup 2 4x 10% 1029   E-424 5  A-16 Butyric none 3x 97%944  E-458 2  A-17 Butyric none 1x 89% 847 E-19 4  E-39 Butyric Workup 21x 85% 819 E-82 4  E-48 Butyric Workup 3 1x 35% 819  E-553 4  E-48Butyric Workup 3 2x 30% 889 E-44 1 A-1 Isobutyric Workup 1 1x 67% 819  E-458, 3  A-17 Butyric Workup 1 1x, 2x, 3x 86%  847,   E-459,  917, E-460 987  E-238 1  A-10 Butyric none 1x 96% 659   E-241, 3  A-10Butyric Workup 1 2x, 3x, 4x 85%  730,   E-249, 800, 870  E-250  E-111 3E-1 Butyric Workup 2 1x 38% 864  E-255* 1  A-11 Butyric Workup 1 2x  3%892  E-256* 1  A-11 Butyric Workup 1 3x 5.6%  964  E-85* 1  A-11 ButyricWorkup 1 3x 964  E-257* 1  A-11 Butyric Workup 1 4x  3% 1036  E-24 3 E-1Butyric Workup 1 1x 75 864 E-89 1 A-3 Butyric n/a 1x  9% 406 *isolatedfrom one reaction

Example 49

Formation of Valeric Esters of ALCs

Method 1. The ALC was taken up in DCM. To this were added pyridine andvaleric acid anhydride (1 equiv. pyridine/1 equiv. valeric acidanhydride). The mixture was stirred at room overnight or over theweekend and was then poured on an aqueous citric acid solution (5% or10%; at RT and was stirred for 15 min. The aqueous phase was extractedwith EtOAc (2×) and was afterwards brought to pH=8 with solid Na₂CO₃.The alkaline aqueous layer was extracted with EtOAc (2×) and thecombined organic phases were washed with water (1×) and saturatedaqueous NaCl-solution (1×), dried (Na₂SO₄), concentrated to dryness anddried at the oil pump. Products were obtained as colorless solids orfoams.

Method 2. Analogue to 2A but after stirring at RT for 2 h additionalpyridine (2 equiv.) and valeric acid anhydride (2 equiv.) were added andstirring was continued overnight. Products were obtained as colorlessfoams or solids.

Method 3. The ALC was taken up in DCM. To this were added pyridine (4equiv.) and valeric acid anhydride (4 equiv.). The mixture was stirredat room overnight or over the weekend. Additional pyridine (2 equiv.)and valeric acid anhydride (2 equiv.) were added and the mixture wasstirred overnight. Reaction mixture was filled into a separation funneland washed with saturated aqueous NH₄Cl-solution (3×) and water (3×).The organic phase was dried (Na₂SO₄) and concentrated to dryness. Theresidue was co-evaporated with toluene (3×) and with DCM (3×).Afterwards the crude product was purified by column chromatography onsilica gel. Eluent: Chloroform/Isopropanol/NH₃ (7 M in Methanol) 30/1/1

The product was dried at the oil pump. Products were obtained ascolorless solids or foams.

TABLE 16 Valeric Ester Examples Overall MS Compound Synthesis equiv. ofacid Degree of m/z Entry Method ALC or anhydride Re Substitution Yield([M + H]⁺) E-72  3 A-1 6 Amount DCM: 5 mL 2 65% 917.5 E-558 1 A 2 5Anhydro erythromycin 1  7% 800.5 (Anhydro) products are formed AmountDCM: 25 mL E-569 3  A-10 6 Amount DCM: 5 mL 2 45% 757.5 E-582 2  A-12 6Amount DCM: 10 mL 1 89% 675.5 Citric acid: 10% E-411 1  A-15 4 AmountDCM; 25 mL 1 and 2 81% 878.3 E-412 Citric acid: 5% 962.3 E-428 1  A-16 3Amount DCM: 25 mL 1 and 2 83% 819.0 E-429 Citric acid: 5% 902.8 E-464 1A17 5 Amount DCM: 25 mL 1 94% 861.7 Citric acid: 5%

Example 50

Formation of Isovaleric Esters of ALCs

Method 1: Isovaleric acid (4.4 equiv./equiv. ALC) and HOBt 85% (4.4equiv./equiv. ALC) were dissolved in DMF (12.5 mml/mmol ALC). Thesolution Was cooled down to 0-5° C. in an ice-bath. At this temperaturea solution of Dicyclohexylcarbodiimide (4.5 equiv./equiv. ALC) in DCM (5ml/mmol ALC) was added dropwise within 30 min. the solution was kept atthis temperature for another 10 min. Then Azithromycin (1 equiv.) wasadded in one portion. While stirring, the solution was allowed to cometo room temperature within 2 h. Stirring was continued for another 2 hat 50° C. The reaction mixture was allowed to stand at RT for 12 h. Awhite precipitate was removed by suction. The solvent was evaporatedcompletely at 12 mbar and 50° C. The residue was dissolved in DCM (12.5mL/mmol ALC) and washed with water (7.5/mmol ALC). A small amount of awhite precipitate was removed. Then the solution was treated with citricacid (25 mL/mmol ALC, 5%). The aqueous phase was washed with DCM (5ml/mmol ALC). NaOH 10% was added until the aqueous phase was basic (pH12) and was washed with DCM (2×10 ml/mmol ALC). After phase separationthe organic phase was evaporated to dryness, products were obtained aswhite solids.

Method 2A. The ALC was taken up in DCM. To this were added pyridine andisovaleric acid anhydride (1 equiv. pyridine/1 equiv. isovaleric acidanhydride). The mixture was stirred at room overnight or over theweekend and was then poured on an aqueous citric acid solution (5% or10%) at RT and was stirred for 15 min. The aqueous phase was extractedwith EtOAc (2×) and was afterwards brought to pH=8 with solid Na₂CO₃.The alkaline aqueous layer was extracted with EtOAc (2×) and thecombined organic phases were washed with water (1×) and saturatedaqueous NaCl-solution (1×), dried (Na₂SO₄), concentrated to dryness anddried at the oil pump. Products were obtained as colorless solids orfoams.

Method 2B. Analogue to 2A but after stirring at room temperature for 2 hadditional pyridine (2 equiv.) and isovaleric acid anhydride (2 equiv.)were added and stirring was continued overnight.

Products were obtained as colorless foams or solids.

Method 2C. The ALC was taken up in DCM. To this were added pyridine (4equiv.) and isovaleric acid anhydride (4 equiv.). The mixture wasstirred at room for approximately 2 h, then a catalytic amount of DMAPwas added, followed by another catalytic amount approximately another 2h later. The mixture was stirred at room temperature for approximately 2h before additional pyridine (2 equiv.) and isovaleric acid anhydride (2equiv.) were added. The mixture was stirred at room overnight and thenpoured on an aqueous citric acid solution (5%,) and stirred at roomtemperature for 30 min. The aqueous phase was extracted with EtOAc andafterwards brought to pH=8 with solid Na₂CO₃. The alkaline aqueous layerwas extracted with EtOAc (2×) and the combined organic phases werewashed with water (1×) and saturated aqueous NaCl-solution (1×), dried(Na₂SO₄), concentrated to dryness and dried at the oil pump. Productswere obtained as colorless solids or foams.

TABLE 17 Isovaleric Ester Examples Overall MS Compound Synthesis equiv.of acid Degree of m/z Entry Method ALC or anhydride AnnotationsSubstitution Yield ([M + H]⁺) E-45  1  A-1 1 91% 833.5 E-557 2A A 2 5Anhydro erythromycin 1 17% 800.5 (Anhydro) products are formed AmountDCM: 25 mL Citric acid: 5% E-573 2C A-10 6 Amount DCM: 7.5 mL 2 19%757.5 Citric acid: 5% E-583 28 A12  6 Amount DCM: 10 mL 1 83% 675.5Citric acid: 10% E-413 2A A-15 4 Amount DCM: 25 mL 1 95% 878.3 Citricacid: 5% E-431 2A A-16 3 Amount DCM: 25 mL 2 87% 818.8 E-432 Citricacid: 5% 902.7 E-467 2A A-17 5 Amount DCM: 25 mL 1 90% 861.6 Citricacid: 5%

Example 51

Long Chain (>C5) Fatty Acid Substation of Tildipirosin

Hexanoic acid (290 mg, 2.5 mmol) was taken up in 5 mL dichloromethane(DCM). Compound A-16 (367 mg, 0.5 mmol) and 4-Dimethylaminopyridine(DMAP) (336 mg, 2.75 mmol) were added and the resulting solution wascooled in an ice bath for approximately 10 minutes. At this point,dicyclohexylcarbodiimide (DCC) (567 mg, 2.75 mmol) was added slowly. Thereaction was stirred continually at this temperature for 5 minutes andthen progressively warmed to room temperature where it was stirredovernight. Dicyclohexylurea (DCU) that was formed during the reaction isfiltered off and discarded. The filtrate was collected and then washedwith a saturated solution of ammonium chloride (3×), sodiumhydrogencarbonate (3×), water (1×) and dried over anhydrous Na₂SO₄. Thesolvent was evaporated in vacuo. This was followed by re-dissolving theresidue in a small volume of methanol. The solution was transporteddropwise into ice-cold water (2× volume of methanol) and stored in thefreezer overnight. The precipitated product was filtered off and driedunder high-vacuum to produce a mixture of E-437, E-438 and E-439 (56%).

This esterification conditions can be extended for other acids.

TABLE 18 Long chain fatty acid substitution of ALC Degree of CompoundSubstituent Substitu- Entry Acid equivalent tion Yield MS E-437,Hexanoic acid 5 2 and 3 56% 931.7 E-438, 1028.8 E-439 E-440, Heptanoicacid 5 1, 2 and 3 46% 846.9 E-441, 959.3 E-442 1071.9 E-445 Octanoicacid 5 3 17% 1113.5 E-447, Decanoic acid 5 2 and 3 41% 1043.3 E-4481197.5 E-450, Dodecanoic 5 2 and 3 60% 1099.5 E-451 acid 1281.4

Example 52

General Procedure for Preparing Cores A-13 and A-14

Macrolide (1 mmol) is dissolved in DMF (500 μl). Epichlorohydrin (1 mL)is added and the mixture is heated to 80° C. for 12 h. When MS analysisindicates complete conversion, all volatiles are removed in vacuo andthe residue is dissolved in ethanol (1 ml). The solution is poured into25 ml of water. The precipitate is isolated and can be used directly forthe next step or is chromatographed to obtain the pure epoxide.

The following macrolides were used to form epoxides as precursor to thedesired cores.

TABLE 19 Epoxide Formation MW Entry Macrolide epoxide Yield 1Azithromycin 703 25% 2 Gamithromycin 731 12% 33-decladinosyl-3-oxoazithromycin 543  5% 4 Tildipirosin 689  5%

Epoxide (1 mmol) is dissolved in 2-propanol (500 μl), and an excess of 5equivalent of an amine is added. The mixture is heated from 12 h to 100h at 80° C. When MS indicates complete conversion, all volatiles areevaporated and the residue subjected to chromatography to separate the 2regioisomeric amines.

TABLE 20 Epoxide opening by amines Epoxide Entry opening (Table 9) Amineposition R¹ R² [M + H]⁺ Yield [%] Comment 1 dimethylamine 3′ Me Me 74966 NMR identical to azithromycin 2′ 22 regioisomer of Azithromycin 1morpholine 3′ R¹ = R² = morpholine ring 791 30 unpolar product (A-13.1)2′ 61 polar product (A-13.2) 1 diethanolamine 3′ C₂H₄OH C₂H₄OH 809 22unpolar product (A-14.1) 2′ 47 polar product (A-14.2) 1 ammonia H H 721n.d.* 1 N-methyl Me OH 751 n.d.* hydroxylamine 1 Iminodiacetic—CH₂C(O)OEt —CH₂C(O)OEt 893 n.d.* some cyclized acid product withdiethylester [M + H]⁺ = 865 is formed, too 2 morpholine R¹ = R² =morpholine ring n.d.* *n.d. not determined

Example 53

Further Acylation Reactions of ALC

Method 1: Compound A-1 (2000 mg, 2.67 mmol) was taken up in 10 mLdichloromethane and stirred. Separately 4.4 eq of a carboxylic acid and4.4 eq of 1,1′-Carbonyldiimidazole were solved in dichloromethane (10mL) and stirred over 20 min. Both solutions were unified and stirredcontinually at room temperature. The dichloromethane phase was washedwith saturated NaHCO₃ solution (2×) and dried with Na₂SO₄ (anhydrous).The solvent was evaporated in vacuo to produce a white foam containingproducts of reaction.

Method 2: Compound E-1 (265 mg, 0.33 mmol) was taken up in 10 mLdichloromethane and stirred. Separately 1.6 eq of methoxyacetic acid and1.6 eq of 1,1′-Carbonyldiimidazole were solved in dichloromethane (5 mL)and stirred over 20 min. Both solutions were unified and stirredcontinually at room temperature. The reaction solution was washed with5% citric acid three times to extract the product. Acidic solution wasthen washed with ethyl acetate (2×) and afterwards neutralized withNa₂CO₃. Product was extracted with ethyl acetate (3×). The solution waswashed with a saturated solution of sodium chloride (2×), water (2×) anddried over anhydrous Na₂SO₄. The solvent was evaporated in vacuo toproduce a white foam containing E12 (238 mg, 90%).

Workup 1: Column chromatography over silica gel was carried out toseparate different products. As eluent a mixture of chloroform,2-propanol and ammonia in methanol (60:1:1) was used. The solvent wasevaporated in vacuo.

Workup 2: Column chromatography over silica gel was carried out toseparate different products. As eluent a mixture of cyclohexane, acetone(3:1) with 0.5% triethylamine was used. The solvent was evaporated invacuo.

TABLE 21 Typical Products from the Acylation Procedures (2) Reaction CpdSynthesis Condition Degree of Entry Method Carboxylic acid ALC (i.e.Workup) Substitution Yield MS E-545 1 Cyclopropanecarboxylic acid A-1Workup 1 1×   30% 817 E-546 1 Cyclobutanecarboxylic acid A-1 Workup 1 1×  10% 830  E-546, 1 Cyclobutanecarboxylic acid A-1 Workup 1 1×, 2×, 3×22.5%  830,  E-560,  913, E-561 995 E-547 1 Nicotinic acid A-1 Workup 21× 10.3% 854  E-547, 1 Nicotinic acid A-1 Workup 2 1×, 2× 11.8%  854,E-562 959 E-551 1 Methoxyacetic acid A-1 Workup 1 1× 4.4 820 E-14,  1Methoxyacetic acid A-1 2×, 3×, 4× Reaction  893, E-22,  solution  965,E-559 1037  E-12  2 Methoxyacetic acid E-1 none 1×   90% 866 E-81  13-Phenylpropionic acid A-1 Workup 1 1×   30% 881 E-544  1*Indole-3-propionic acid A-1 Workup 1 1×   45% 920 *instead of1,1′-carbonyl diimidazole, HATU was used as coupling agent.

Example 54

Synthesis of E-541 and E-542

E-27 (1.2 mmol) and Quinoline-amine (1 eq) was taken up in DCM (5 mL).To this was added HATU (1.2 eq) neat. The reaction was stirred overnightat room temperature. Reaction was very sluggish an additional 0.5 eq ofHATU was added. Reaction was stirred for 2 days or disappearance ofstarting material was observed (TLC or MS). Reaction solution wasremoved in vacuo and the crude material directly purified bychromatography to get the desired product.

TABLE 22 Decoration of ALC (E-27) with potential TLR-agents Cpd Degreeof Entry Quinoline Substitution Yield MS E-541 Imiquimod 1 35% 1071E-542 Resiquimod 1 25% 1145

Example 55

Examples of Polyamines as ALC

These ALCs can be prepared by reacting symmetrical or unsymmetrical di-or poly-epoxides with secondary amines. This will provide ALCs thatcontains common structural element of 2 or more alcohols, vicinallyneighbored by a tertiary amine. Some polyamines are also commerciallyavailable.

Alternatively, ALCs can be prepared by reacting epoxides withdiethanolamine. The reaction products are containing 2-hydroxy tertiaryamines.

General Procedure for Preparation of some polyamine ALC Bearing hydroxyFunctionalities:

Polyamine (1 mmol) containing at least 2 NH-functions and the epoxideare mixed and heated without solvent to 80° C. Excess epoxide can beremoved by column chromatography selectively. Products are sufficient,when at least 2 tertiary ß-hydroxyamines are present.

TABLE 23 Examples of Polyamine ALCs Eq. of Reaction [M + H]⁺ Entry AmineEpoxide Epoxide time Product Yield Remark 1 hexamethylene diamine 1,2-4.2  40 h 966 n.d. product contains epoxytetradecane a small amount oftriple alkylation 2 bis- cyclohexene 12 240 h 608 n.d. main product is(hexamethylene)triamine oxide tetraalkylated

Reactions of Polyepoxides with Amines:

Corresponding polyepoxide (1 mmol) is mixed with 1.05 mmol secondaryamine per epoxide function and heated to 80° C. without solvent for 12h.

TABLE 24 Further Examples Polyamine ALCs [M + H]⁺ Entry Epoxide AmineProduct Yield 1 1,2,7,8-diepoxyoctane morpholine 317 100% 2 Diglycidylethylenglycol morpholine 377  90%

Synthesis of E-552

1.45 g of 1,10-dimorpholino-2,9-dihydoxy-4,7-dioxadecane are combinedwith 1.5 ml of butyric anhydride in 5 ml of chloroform. After stirringfor 1 h, MS indicates complete conversion ([M+H]⁺=517). The mixture isextracted with 2 N KOH, saturated aqueous sodium bicarbonate solutionand brine, dried and chromatographed over silica gel to obtain 1.57 g ofthe target compound (72%).

Example 56

Substitution of Polyamine ALC (see Table 1, Entry A-19)

Substitution ALC A-19.1/A-20.1/A-20.2

Method 1: N-Hydroxyalkyl compound (5 mmol) was suspended in excesscarboxyl acid anhydride (>100 mmol, >20 eq.) in a round bottom flaskwhile stirring (magnetic stir bar, 500 rpm). The mixture was cooled inan ice bath and sulfuric acid (>96%, 3 drops) was carefully added ascatalyst. Stirring was continued until a clear solution was obtained.When ESI-MS indicated full conversion of starting materials the reactionmixture was poured on ice. The system was stirred for 2 or more hours inorder to hydrolyze any anhydride. The mixture was neutralized byaddition of sodium bicarbonate and extracted with dichloromethane (3×).Separation of organic phase, drying over sodium sulfate and evaporationof any volatiles in vacuo yielded the product as colorless oil.

Method 2. Carboxylic acid (1.5 eq per hydroxyl group) was placed into around bottom flask along with a stir bar and carbonyl diimidazole (CDI,1.6 eq per hydroxyl group). Dichloromethane (DCM, 10 mL per gramcarboxylic acid) was added carefully while stirring (500 rpm) at ambienttemperature. Immediate formation of carbon dioxide indicated conversionof corresponding acid to the acyl donor (caution: too quick CO₂formation may result in strong foaming. Do not seal the flask!). After acouple of minutes, a clear solution was obtained and stirring wascontinued for 15 minutes. N-Hydroxyalkyl compound (5 mmol) was added atambient temperature and the reaction mixture was stirred overnight. WhenESI-MS indicated full conversion of starting materials the reaction wasquenched by addition of methanol, converting excess acyl donor to methylester. The system was diluted by addition of further DCM and was subjectto extraction with saturated sodium bicarbonate solution (3×).Separation of organic phase, drying over sodium sulfate and evaporationof any volatiles in vacuo yielded the product as colorless oil.

TABLE 25 Substitution of ALC A-19.1/A-20.1/A-20.2 Compound SynthesisDegree of MS Entry Method ALC Substitution Yield [M + H⁺ E-524 1 A-19.1Di acyl 85% 259, M + H⁺ E-576 1 A-19.1 Mono acyl 23% 217, M + H⁺ E-525 1A-19.1 Di acyl 94% 287, M + H⁺ E-577 1 A-19.1 Mono acyl 12% 231, M + H⁺E-526 2 A-19.1 Di acyl 87% 315, M + H⁺ E-578 2 A-19.1 Mono acyl 45% 245,M + H⁺ E-527 1 A-19.1 Di acyl 56% 315, M + H⁺ E-529 1 A-19.1 Di acyl  8%343, M + H⁺ E-530 2 A-19.1 Di acyl 76% 499, M + H⁺ E-538 1 A-20.1 Triacyl 85% 299, M + Na⁺ E-537 1 A-20.1 Tri acyl 76% 383, M + Na⁺ E-580 1A-20.2 Tri acyl 61% 391, M + Na⁺; 368, M⁻

Substitution ALC A-19.2

1,1′-Carbonyldiimidazole was dissolved in dichloromethane (dry, 25 mL)and to this was added the carboxylic acid slowly at room temperature.The solution was stirred at room temperature before a suspension ofN;N,N′,N′-tetrakis (2-hydroxyethyl)-ethylendiamine (A-19.2) indichloromethane (dry, 5 mL) was added in one portion at room temperatureand the mixture was stirred at RT. The reaction mixture was filled intoa separation funnel and washed. The organic phase was dried (Na₂SO₄) andconcentrated to dryness in vacuo. The crude product was purified bycolumn chromatography.

Eluent: CHCl₃: Isopropanol: NH₃ (7 M in MeOH)=60:1:1.

TABLE 26 Substitution of ALC A-19.12 Amount Time for Carboxylic stirringCpd Amount acid Amount carboxylic Reaction Washing Entry ALC (A-19.2)(mg) CDI acid and CDI Time steps Yield E-531 A-19.2 521 mg Glacialacetic 2.49 g 35 min 22h 30 min NaHCO₃ 297 mg (73.4%) acid 15.35 (2 × 20(45%) 1.64 800 μL mmol mL) mmol 13.99 mmol E-532 A-19.2 498 mg Propionicacid 2.46 g 20 min 1.5 h with Water 144 mg (73.4%) 943 μL 15.17 withargon argon stream (1 × 20 (20%) 1.55 12.6 mmol mmol stream 69 h 45 minmL) mmol under argon sat. atmosphere NaHCO₃ (2 × 20 mL) E-533 A-19.2 495mg Butyric acid 2.51 g 35 min 46 h 45 min NaHCO₃ 20 mg (73.4%) 1.2 mL15.45 (2 × 20 (pure) 1.54 13.55 mmol mmol mL) 224 mg mmol (with impuri-Ties Overall yield: (19%)

Analytical Data:

E-531:

ESI-MS (positive): m/z=405.2 [M+H]⁺, 427.1 [M+Na]⁺

Purity according to HPLC (ELSD): >99.9%

¹H-NMR (300 MHz, CDCl₃): 1.98 (s, 12 H, 4-H, 4′-H, 4″-H, 4″′-H), 2.57(s, 4 H, 1-H, 1′H), 2.72 (t, J_(2,3) and J_(2′,3′), J_(2″,3″),J_(2″′,3″′)=6.04, 8 H, 2-H, 2′-H, 2″-H, 2″′-H), 2.72 (t, J_(3,2) andJ_(3′,2′), J_(3″,2″), J_(3″′,2″′)=4.04, 8 H, 2-H, 2′-H, 2″-H, 2″′-H).

¹³C-NMR (75 MHz, CDCl₃): 20.77 (q, C-4, C-4′, C-4″, C-4″′), 53.15 (t,C-2, C-2′, C-2″, C-2″′), 53.44 (t, C-1, C-1′), 62.43 (t, C-3, C-3′,C-3″, C-3″′), 170.74 (s, 4×C=O).

E-532:

ESI-MS (positive): m/z=461.2 [M+H]⁺, 483.3 [M+Na]⁺

Purity according to HPLC (ELSD): >99.9%

¹H-NMR (300 MHz, CDCl₃): 1.08 (t, J_(5,4), J_(5′,4′), J_(5″,4″),J_(5″′,4″′)=7.6 Hz, 12 H, 5-H, 5′-H, 5″-H, 5″40 -H), 2.27 (q, J_(4,5),J_(4′,5′), J_(4″,5″), J_(4″′,5″′)=7.6 Hz), 2.57, 4-H, 4′-H, 4″-H, 4 ″′2.60 (s, 4 H, 1-H, 1′H), 2.74 (t, J_(2,3) and J_(2′,3′), J_(2″,3″),J_(2″′,3″′)=6.04, 8 H, 2-H, 2′-H, 241 -H, 2″′-H), 4.07 (t, J_(3,2) andJ_(3′,2′), J_(3″,2″), J_(3″′)=6.04, 8 H, 2-H, 2′-H, 2″-H, 2″′-H).

¹³C-NMR (75 MHz, CDCl₃): 8.96 (q, C-5, C-5′, C-5″, C-5″′), 27.44 (t,C-4, C-4′, C-4″, C-4″′), 53.22 (t, C-2, C-2′, C-2″, C-2″′), 53.54 (t,C-1, C-1′), 62.36 (t, C-3, C-3′, C-3″, C-3″′), 172.20 (s, 4×C=O).

E-533:

ESI-MS (positive): m/z=617.3 [M+H]⁺, 539.3 [M+Na]⁺

Purity according to HPLC (ELSD): >99.9%

¹³C-NMR (75 MHz, CDCl₃): 13.53 (q, C-6, C-6′, C-6″, C-6″′) 18.26 (t,C-5, C-5′, C-5″, C-5″′), 36.00 (t, C-4, C-4′, C-4″, C-4″′), 53.21 (t,C-2, C-2′, C-2″, C-2″′), 53.45 (t, C-1, C-1′), 62.24 (t, C-3, C-3′,C-3″, C-3″′), 173.36 (s, 4×C=O):

Substitution ALC A-19.3

H-L-orn(Boc)2CT Resin (0.68 mmol/g, 100-200 mesh, 2.99 g, 2.07 mmol) wasfilled into a 20 mL syringe with frit. Dichloromethane (dry, 10 mL),MeOH (2 mL) and diisopropylethylamine (2 mL) are added to the resin forendcapping. The mixture was shaken at room temperature for 30 min, thenthe liquid was sucked off and the resin was washed (3× dimethylformamide15 mL, 1× diethylether 15 mL).

The resin was filled into a 100 mL round bottom flask. DMF (25 mL) wasadded and the resin was swollen for 5 min. Then diisopropylethylamine(3.8 mL, 22.3 mmol) and 2-bromoethanol (1.434 mL, 20.3 mmol) were addedsubsequently at room temperature. The reaction mixture was stirred at60° C. (bath temperature) for 24 h.

The resin was filled into a 20 mL syringe with frit and was washed:

4× dimethylformamide (20 mL), 3× methanol (20 mL), 3× dichloromethane(20 mL), 3× diethyl ether (20 mL).

Half of the resin (1.035 mmol) was filled into a 20 mL syringe withfrit.

Valeric acid (568 μL, 5.69 mmol) was added to a mixture ofdimethylformamide/dichloro-methane 1:1 (10 mL). HOBt*H₂O (870 mg, 5.69mmol) was added and mixture was stirred at room temperature for 5 minbefore diisipropylcarbodiimide (881 μL, 5.69 mmol) was added. Stirringat room temperature was continued for 10 min, then the whole mixture wasadded to the resin and the resin was shaken at room temperature for 5 h.

The liquid was sucked off and the resin was washed:

4× dimethylformamide (10 mL), 3× methanol (10 mL), 3× dichloromethane(10 mL), 3× diethyl ether (10 mL).

A test cleavage showed the product by mass spectrometry

ESI-MS (positive): m/z=261.1 [M+H]⁺

Example 57

Synthesis of 2′-O-(2-Ferrocenyl) Acetyl-Azithromycin E-549

ALC A-1 (0.25 mmol) was dissolved in dry dichloromethane (5 mL). To thiswas added EDCI (2 eq., 0.5 mmol) and 2-ferrocenyl acetic acid (1.1 eq.,0.28 mmol). The reaction was stirred overnight at room temperature. Thesolvent was removed in vacuo and the resulting white amorphous foam. Theresulting crude product was purified by column chromatography with agradient starting at 10% of acetone in cyclohexane (0.2% Et₃N).

Example 58

Synthesis of E-258

Compound A-11/E-16 was dissolved in dry dichloromethane (DCM) in a roundbottom flask equipped with magnetic stir bar.Penta-O-acetyl-α-D-mannopyranoside (1.2 eq) was added and the system wascooled in an ice bath while stirring (300 rpm). Catalytic amount ofboron trifluoride diethyl ether complex was carefully added dropwise andthe system was allowed to warm up while stirring overnight. Upondilution with further DCM the mixture was subject to extraction withsaturated sodium bicarbonate solution (3×). Separation of organic phase,drying over sodium sulfate and evaporation of any volatiles in vacuoyielded the product as colorless oil or beige to off-white foam. [M+H]⁺m/z 907

The reaction conditions also produced the des-cladinosyl product E-600.

Example 59

Synthesis of E-550

350 mg of compound E 77 are dissolved with 15 ml of carbon disulfide.250 mg of sulfur are added and the mixture is stirred for 7 days. Themixture is extracted with 5% aqueous citric acid solution. The aqueousextract is combined with 10 ml of ethyl acetate and made alkaline byaddition of sodium carbonate with intense stirring. The organic phase isseparated, washed with brine, dried over sodium sulfate and concentratedin vacuo to yield 280 mg of a product, that contains various highersulfides along with some starting material, as indicated by massspectrometry ([M+H]⁺=969, 1001, 1033, 1065).

Example 60

Pharmacokinetics

The distribution of compounds to target organs is of specific importanceto the efficacy of anti-infective compounds. To determine distributionthe compounds are formulated and administered to a suitable animalmodel. Compounds were administered p.o. 10 mg/kg in 2% citric acid inBALBc and organs were recovered at 6 h. Organs were extracted inAcetonitrile (6× volume of the sample), centrifuged at 14000 g for 5minutes. Samples were analysed by LCMSMS (SCIEX 4500). Data are the meanof 3 animals.

Example 61

Selection of ALC via Concentration into Immune Cells

The distribution of compounds to target cells is of specific importanceto the efficacy of anti-infective compounds. To determine uptake thecompounds are dissolved in DMSO or citric acid and mixed with wholeblood, plasma or cell medium. To these solutions are added culturedmacrophages, cultured immune cells, bone marrow derived macrophages,peritoneal macrophages or buffy coat cells. The mixture is incubated at37° C. for 1, 2, or 3 hours. After incubation, the immune cells areseparated from the medium and the concentration of the compounds isdetermined by extraction in Acetonitrile (6× volume of the sample),followed by centrifugation at 14000 g for 5 minutes. The resultingextracts are analyzed by LCMSMS (SCIEX 4500 in positive mode). Data arethe mean of 3 animals.

Other Embodiments

All of the features disclosed in this specification may be combined inany combination. Thus, unless expressly stated otherwise, each featuredisclosed is only an example of a generic series of equivalent orsimilar features.

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

Abbreviations

The following abbreviations were used as noted:

-   -   MeOH: methanol    -   NaHCO₃: sodium bicarbonate    -   K₂CO₃: potassium carbonate    -   MS: mass spectrometry    -   DMSO: dimethyl sulfoxide    -   TLC: thinlayer chromatography    -   Et₃N: triethylamine    -   EtOAc: ethyl acetate    -   DCM: dichloromethane    -   NH₄Cl: ammonium chloride    -   THF: tetrahydrofuran    -   Na₂CO₃: sodium carbonate    -   EDCI: N-Ethyl-N′-(3-dimethylaminopropyl)carbodiimide        hydrochloride    -   DMAP: 4-dimethylamino pyridine    -   HATU        O-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-hexafluorphosphat    -   DIPEA N,N-Diisopropylethylamine

CITATION LIST Patent Literature

US2007238882A1

US2003105066A1

US2008221158A1

US2008027012A1

US6455576B1

US5677287A

EP1748994B1

WO2007025632A2

WO9530641A1

WO0002567A1

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1. A compound, or salt thereof, comprising an amphiphilic lysosomallytrapped compound (alc) conjugated via an ester, thioester, amide, ornitroester to one or more products of anaerobic metabolism (pams) of thesame or different types.
 2. The compound as in claim 1 in which the PAMis selected from one or more of Short Chain Fatty Acid (SCFA), NO, H₂S,mercaptans, polyamines, decarboxylated amino acids, TLR ligands, orpolyphenol metabolites like phenylpropionic acid.
 3. The compound as inclaim 1 in which the ALC is selected from a macrolide, polyamine,propranolol analog, chloroquine analog, amodiaquine, dextromethorphan,dextrorphan, paroxetine, fluoxetine, astemizole or imipramine analog. 4.A method of stimulating immune or epithelial cells to form ananti-infective barrier or anti-infective response comprising contactingthe cells with a compound of Formula 1, or salt thereof, wherein thecontacting results in the intracellular release of a PAM comprising oneor more of a molecule type selected from TLR ligands, SCFA, NO, H₂S,sulfides, polyamines, decarboxylated amino acids or polyphenolmetabolites like phenylpropionic acid from the compound of Formula 1, orsalt thereof.
 5. The method as in claim 4 comprising the intracellularrelease of one or more types of a short chain fatty acid moiety from thecompound of Formula
 1. 6. The method as in claim 5 comprising theintracellular release of a short chain fatty acid moiety containing 2 ormore carbons from an appropriate carrier molecule.
 7. (canceled)
 8. Thecompound of claim 1, wherein the compound is one of the following:(Formula 2)

Wherein, X=—N(CH₃)—CH₂—; —CH₂—N[(CH₂)_(n)—CH₃]— wherein n is 0-4;—C(═O)—; —C(═NOR⁸)—; —C(═NR¹²)—; R₁ is: -(C₁-C₁₀)alkyl;-(C₁-C₁₀)alkyliden-OH; -(C₁-C₁₀)alkyliden-ONO₂; R₂ is: —H; —NO_((y))with y=1 or 2; —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷,—C(═S)(NH)R⁷; R³ is: —H; —NO_((y)) with y=1 or 2; —C(═O)OR⁷, —C(═S)OR⁷,—C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷, —C(═S)(NH)R⁷; If Z=O, R₄ is: —H;—NO_((y)) with y=1 or 2; —C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷,—C(═O)(NH)R⁷, —C(═S)(NH)R⁷; R⁵ is: —H; —NO_((y)) with y=1 or 2;—C(═O)OR⁷, —C(═S)OR⁷, —C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷, —C(═S)(NH)R⁷; orZ=O or NR⁹ and the R₄ and R⁵ bearing atoms are connected via —C(═O)— (IfZ=O: carbonate linkage, if Z=NR⁹: carbamate linkage); or the R₄ and R⁵bearing atoms are connected via W; W is: —(—)CH-(C₁-C₁₂)alkyl;—(—)CH-(C₃-C₁₂)alkenyl; —(—)CH-(C₃-C₁₂)alkynyl;—(—)CH-(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;—(—)CH-(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl; wherein alkyl, alkenyl, alkynylare optionally substituted by one to five substituents selectedindependently from halogen, (C₁-C₄)alkyl, (C₁-C₄)alkenyl,(C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl, (C₁-C₆)heterocycloalkyl,(C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy, hydroxy, nitro, cyano,azido, mercapto, —NR¹⁴R¹⁵, R¹⁴C(═O)—, R¹⁴C(═O)O—R¹⁴OC(═O)O—,R¹⁴NHC(═O)—, R¹⁴C(═O)NH—, R¹⁴R¹⁵NC(═O)—, R¹⁴OC(═O)—, and —xNO₂ withx=O;S;N; R⁶ is: —H; —NO_((y)) with y=1 or 2; —C(═O)OR⁷, —C(═S)OR⁷,—C(═O)R⁷, —C(═S)R⁷, —C(═O)(NH)R⁷, —C(═S)(NH)R⁷; each R⁷ isindependently: —H; -ferrocene; -C₁-C₁₀ alkyl, alkenyl, alkynyl, aryl,heteroaryl, alkylaryl, alkylheteroaryl wherein alkyl, alkenyl, alkynyl,aryl, heteroaryl, alkylaryl and alkylheteroaryl groups are optionallysubstituted by one to five substituents selected independently from:ferrocene, halogen (as can be F, Cl, Br, I), (C₁-C₄)alkyl,(C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,(C₁-C₆)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy,hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido (—N₃), mercapto (—SH),—NR¹⁴R¹⁵, R¹⁴C(═O)—, R¹⁴C(═O)O—, R¹⁴OC(═O)O—, R¹⁴NHC(═O)—, R¹⁴C(═O)NH—,R¹⁴R¹⁵NC(═O)—, R¹⁴OC(═O)—, and —XNO_((y)) with X=O; S; N and y=1 or 2;R⁸ is: —H; —NO_((y)) with y=1 or 2; —C(═O)—R⁷; -(C₁-C₁₂)alkyl;-(C₁-C₁₂)alkenyl; -(C₁-C₁₂)alkynyl; -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;-(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl; -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;-(C₂-C₉)heteroaryl-(C₁-C₅)alkyl; R⁹ is: —H; —NO_((y)) with y=1 or 2;—C(═O)—R⁷; -(C₁-C₁₂)alkyl; -(C₁-C₁₂)alkenyl; -(C₁-C₁₂)alkynyl;-(C₁-C₈)[(C₁-C₄)alkoxy]alkyl; -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;-(C₆-C₁₀)aryl-(C₁-C₅)alkyl; -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl; R¹² is: —H;—NO_((y)) with y=1 or 2; —C(═O)—R⁷; -(C₁-C₁₂)alkyl; -(C₁-C₁₂)alkenyl;-(C₁-C₁₂)alkynyl; -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;-(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl; -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;-(C₂-C₉)heteroaryl-(C₁-C₅)alkyl; R¹⁴, R¹⁵ are each independently: —H;-(C₁-C₁₂)alkyl; -(C₁-C₁₂)alkenyl; -(C₁-C₁₂)alkynyl;-(C₁-C₈)[(C₁-C₄)alkoxy]alkyl; -(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl;-(C₆-C₁₀)aryl-(C₁-C₅)alkyl; -(C₂-C₉)heteroaryl-(C₁-C₅)alkyl; whereinalkyl, alkenyl, alkynyl, aryl and heteroaryl are optionally substitutedby one to five substituents selected independently from halogen (as canbe F, Cl, Br, I), (C₁-C₄)alkyl, (C₁-C₄)alkenyl, (C₁-C₄)alkynyl,(C₃-C₇)cycloalkyl, (C₁-C₆)heterocycloalkyl, (C₆-C₁₀)aryl,(C₁-C₉)heteroaryl, (C₁-C₄)alkoxy, hydroxyl (—OH), nitro (—NO₂), cyano(—CN), azido (—N₃), mercapto (—SH), and —XNO_(y) with X=O; S; N and y=1or 2; or N(R¹⁴R¹⁵) is an aziridine, azetidine, pyrrolidine, piperidine,azepane or azocane, 1-substituted piperazine, or morpholine moiety. 9.The compound of claim 1, wherein the compound is one of thefollowing:
 1. Formula 3:

Wherein R¹,R²,R⁴,R⁵,R⁶, X and Z are defined as in formula 2; R^(3a),R^(3b)=both —H; or if R^(3a) is —H, then R^(3b) is: —OH; —OR¹⁴; NR¹⁴R¹⁵;—C(═O)—R⁷; or R^(3a)=R^(3b)=(═O); =a cyclic or non-cyclic acetal;(=NR¹²); =a cyclic or non-cyclic aminal; —OC(═O)R⁷; OR¹⁴; and R⁷, R¹⁴,and R¹⁵ are defined as in formula 2 in claim 8; or salt thereof.
 10. Acompound as in:

Where X is O or S; When X=O, R₁ is —(C═O)CH₃, —(C═O)CH₂CH₃,—(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, —(C═O)CHCHCOOH,—(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₂Y, or —(C═O)CH(ONO₂)CH₃; R₂=R₃=H; Y=a 5-memberedsaturated ring containing a disulfide bond; or When X=O, R₁ is—(C═O)CH₃, —(C═O)CH₂CH₃, —(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH,—(C═O)(C═O)CH₃, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₂Y, or—(C═O)CH(ONO₂)CH₃; R₂=CH₃; R3=H; or When X=O, R₁=NO₂; R₂ consists oflinker —CH₂CH₂OR₄, where R₄ is —(C═O)CH₃, —(C═O)CH₂CH₃, —(C═O)CH₂CH₂CH₃,—(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH₃,—(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂, or—(C═O)CH₂CH₂CH₂CH₂Y; R₃=H; Y =a 5-membered saturated ring containing adisulfide bond; or When X=O, R₁ is —(C═O)CH₃, —(C═O)CH₂CH₃,—(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH, —(C═O)(C═O)CH₃, —(C═O)CHCHCOOH,—(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₂Y, or —(C═O)CH(ONO₂)CH₃; Y=a 5-membered saturated ringcontaining a disulfide bond; R₂ consists of linker —CH₂CH₂OR₄, whereR₄=NO₂; R₃=H; or When X=O, R₁ is NO₂, R₂=H or CH₃, R³=OR₅, where R₅ is—(C═O)CH₃, —(C═O)CH₂CH₃, —(C═O)CH₂CH₂CH₃, —(C═O)CH₂CH₂COOH,—(C═O)(C═O)CH₃, —(C═O)CHCHCOOH, —(C═O)CH(OH)CH₃, —(C═O)C(CH₃)₂,—(C═O)CH₂CH₂CH₂CH₃, —(C═O)CH₂C(CH₃)₂, —(C═O)CH₂CH₂CH₂CH₂Y, or—(C═O)CH(ONO₂)CH₃; Y=a 5-membered saturated ring containing a disulfidebond; or When X=S, R₁ is —(C═O)CH₃, a metal salt, or forms a disulfidebridge with itself, R₂=R₃=H.
 11. The compound of claim 3, wherein theALC is selected from a compound of Formula 5:

Wherein Mac=a macrolide ring or macrolide ring system, for example, butnot limited to azithromycin or gamithromycin, each without the desosaminresidue.
 12. The compound of claim 3, wherein the ALC is selected from acompound of Formula 6:

Wherein Mac=a macrolide ring or macrolide ring system, for example, butnot limited to azithromycin or gamithromycin, each without the desosaminresidue R′=independently of each other —H; —NO_((y)) with y=1 or 2;—C(═O)OR³, —C(═S)OR³, —C(═O)R³, —C(═S)R³, —C(═O)(NH)R³, —C(═S)(NH)R³;R¹, R₂=independently of each other H, OH, OR₄, -C₁-C₁₀ alkyl, alkenyl,alkynyl, aryl, heteroaryl, alkylaryl, alkylheteroaryl; wherein alkyl,alkenyl, alkynyl, aryl, heteroaryl, alkylaryl and alkylheteroaryl groupsare optionally substituted by one to five substituents selectedindependently from: fluorine, (C₁-C₄)alkyl, (C₁-C₄)alkenyl,(C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl, (C₁-C₉)heterocycloalkyl,(C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy, hydroxyl (—OH), nitro(—NO₂), cyano (—CN), azido (—N₃), mercapto (—SH), (C₁-C₄)alkthio,—NR⁴R⁵, R⁴C(═O)—, R⁴C(═O)O—, R⁴OC(═O)O—, R⁴NHC(═O)—, R⁴C(═O)NH—,R⁴R⁵NC(═O)—, R⁴OC(═O)—, and —XNO_((y)) with X=O; S; N and y=1 or 2; orN(R¹R²) is an aziridine, azetidine, pyrrolidine, piperidine, azepane orazocane, 1-substituted piperazine, or morpholine moiety; R³=-C₁-C₁₀alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl, alkylheteroarylwherein alkyl, alkenyl, alkynyl, aryl, heteroaryl, alkylaryl andalkylheteroaryl groups are optionally substituted by one to fivesubstituents selected independently from: halogen, (C₁-C₄)alkyl,(C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,(C₁-C₉)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy,hydroxyl (—OH), nitro (—NO₂), cyano (—CN), azido (—N₃), mercapto (—SH),(C₁-C₄)alkthio, —NR⁶R⁷, R⁶C(═O)—, R⁶C(═O)O—, R⁶OC(═O)O—, R⁶NHC(═O)—,R⁶C(═O)NH—, R⁶R⁷NC(═O)—, R⁶OC(═O)—, and —XNO_((y)) with X=O; S; N andy=1 or 2; R⁴, R⁵, R⁶ and R⁷ are each independently: —H; -(C₁-C₁₂)alkyl;-(C₁-C₁₂)alkenyl; -(C₁-C₁₂)akynyl; -(C₁-C₈)[(C₁-C₄)alkoxy]alkyl;-(C₁-C₈)[(C₁-C₄)alkoxy]alkenyl; -(C₆-C₁₀)aryl-(C₁-C₅)alkyl;-(C₂-C₉)heteroaryl-(C₁-C₅)alkyl; wherein alkyl, alkenyl, alkynyl, aryland heteroaryl are optionally substituted by one to five substituentsselected independently from ferrocene, halogen, (C₁-C₄)alkyl,(C₁-C₄)alkenyl, (C₁-C₄)alkynyl, (C₃-C₇)cycloalkyl,(C₁-C₉)heterocycloalkyl, (C₆-C₁₀)aryl, (C₁-C₉)heteroaryl, (C₁-C₄)alkoxy,hydroxyl (—OH), (C₁-C₆)acyloxy, nitro (—NO₂), cyano (—CN), azido (—N₃),mercapto (—SH), and —XNO_(y) with X=O; S; N and y=1 or
 2. 13. Thecompound of claim 8, wherein the number of PAMs is
 0. 14. The compoundof claim 9, wherein the number of PAMs is
 0. 15. A non-nitrate salt of acompound as in claim
 1. 16. A pharmaceutical composition comprising acompound of claim 1, or salt, solvate, or hydrate thereof, and apharmaceutically acceptable carrier.
 17. The composition of claim 16,further comprising an additional therapeutic agent.
 18. A method oftreating a disease, disorder, or symptom thereof in a subject comprisingadministering to the subject a compound of claim 1, or salt, solvate, orhydrate thereof.
 19. The method of claim 18, wherein the disease ordisorder is an infectious disease, an inflammatory disease, or amalignant disease.
 20. The method of claim 18, further comprisingadministering an antibacterial compound.
 21. The method of claim 18,further comprising administering a macrolide compound.